Method of producing a safe, whole-extract of curcuma for oral and topical use

ABSTRACT

Methods of making safe extracts of  Curcuma longa  L. are provided. The processes provided include methods that use an extraction solvent that is at least substantially non-toxic and useful also as a pharmaceutically acceptable carrier in liquid dosage forms. The processes can produce a significantly higher yield from a single extraction than the state-of-the-art processes. For example, liquid dosage forms can be produced directly from the extraction process without requiring removal of the extraction solvent, reducing complexity and cost of processing over the state-of-the-art. Methods of making microemulsions and nanoemulsions are also provided to enhance the bioavailability and stability of the extracts.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.13/590,188, filed Aug. 21, 2012, which is hereby incorporated herein byreference in its entirety.

BACKGROUND

1. Field of the Invention

The teachings provided herein generally relate to formulations and usesof extracts of Curcuma longa L. plants for safe use topically, orally,rectally, or vaginally, for example.

2. Description of Related Art

A New Use of Curcuma longa L. Extract as a Sunscreen

The teachings provided herein shows that the Curcuma longa L. extract,surprisingly, has very good ultraviolet energy absorption propertiesacross the entire wavelength range of interest, making it veryattractive as a sunscreen agent. Moreover, the constituents of Curcumalonga L. have been shown to provide other therapeutic benefits. Forexample, antioxidant and anti-inflammatory effects have been shown, andtreatments for Alzheimer's disease, Parkinson's disease, cysticfibrosis, liver injury, alcohol-induced liver disease, multiplesclerosis, human immunodeficiency virus (HIV), and cancer have beenproposed. As such, the use of Curcuma longa L. extract as a sunscreenagent has the potential of providing additional therapeutic benefitsthat could far exceed merely protecting the skin from the UV rays of thesun.

There are clearly many sunscreen products on the market, and all utilizechemical agents to absorb UV radiation. UV radiation is the radiationthat has shorter wavelengths than visible light and ranges from about 10nm to 400 nm. The lower wavelengths, up to about 290 nm, are screenedout by the earth's ozone layer and are generally of less interest. Assuch, sunscreen products focus on wavelengths ranging from 290 nm to 400nm. Wavelengths ranging from 290 nm to 320 nm are considered “UVB” rays,and although UVB rays have a higher intensity, they also have lesspenetration power. As such, UVB rays may damage the superficial layersof skin but do not penetrate deeper, they do not penetrate glass, andtheir effects can generally be felt greatest between the hours of 10 aand 4 p in the US. Wavelengths ranging from 320 nm to 400 nm areconsidered “UVA” rays. UVA is less intense but penetrates deeper intothe skin to cause damage to the deeper layers, through glass, and itseffects are not limited to certain hours of the day. UVA has beenconsidered to be less damaging than UVB due to it's lower intensity, andbecause UVB light has been identified as primarily responsible forsunburn, as well as melanoma and other skin cancers. However, UVA/sability to cause damage at deeper layers of the skin, and because it'sabout 30 to 50 times more prevalent than UVB, much attention hasrecently been focused on UVA radiation. It has been shown, for example,that UVA can potentiate carcinogenesis from UVB and affects immunefunction. Interestingly, on Jun. 17, 2011, the FDA reported thatsunscreen products should have both UVA and UVB protection, and that theUVA range has two unique areas of interest, the UVA₁ area ranging from340 nm to 400 nm and the UVA₂ area ranging from 320 nm to 340 nm. TheFDA stated that too much emphasis has been placed on UVB, that at least20% of the protection should be in the UVA₂ region, and at least 60% ofthe protection should be in the UVA₁ region, stressing that the UVA₁region is important.

A problem is that currently available sunscreen chemicals absorb lightin either the UVA or UVB range. As such, the desired level of protectionfor the consumer, as indicated by at least the FDA, is not readilyavailable. Currently, to address this problem, the art uses (i) titaniumdioxide to extend the UVA range of protection, (ii) a combination ofsunscreen agents to overlap and broaden the spectrum of UV protection,or (iii) a combination of titanium dioxide and overlapping ranges.Significant problems still remain, however. One problem is that theoverlapping of peaks leaves skin exposed to weak protection in areas ofthe UV spectrum due to an inability to adequately overlap protectionacross the UV spectrum. Another problem is that the titanium dioxide canpenetrate the skin, and this makes it particularly problematic in thatit can potentially convert to compounds that pose a cancer risk whenexposed to the UV energy.

The Problems of Producing a Curcuma longa L. Extract

Extraction methods that currently exist suffer several problems. Oneproblem is that these methods incorporate undesirable chemicals atundesirable levels, for example, ethylene dichloride, methylenedichloride, and ethyl acetate. Ethanol can also be used, but it can onlybe present to a limited extent for internal consumption, and topicalapplication is also limited due to side effects. As such, much efforthas been spent trying to keep residual solvents below limits. Currentextraction processes, for example, generally require complex processesthat include removal of undesirable or toxic extraction solvents to meetFDA rules, and this can include, for example, distillation of thesolvent to form a powder of the extract, a back-extraction of theextract into a different solvent that is suitable for a particular use,complexing with metal ions, use of high pH to precipitate a complex withammonia, washing with methanol and water, and drying at hightemperatures. In view of at least the above, current extraction methodssuffer from (i) use of toxic solvents that require steps to remove andrisk the presence of toxic residuals; (ii) heating in multiple steps toextract and purify; (iii) use of a high pH up to 9.5 on Curcuma longaL., which degrades above pH 7.0; and, ipso facto (iv) multi-stepprocesses that are complex and expensive. There is currently noextraction process for Curcuma longa L. that uses a solvent that issuitable for both topical use and internal consumption.

Improving the Use of Curcuma longa L. Extract in Other Administrations,Such as Oral or Rectal Administrations

Herbal preparations, including Curcuma root, are offered commercially,usually as capsules containing the dried and ground plant material.Curcuma longa L. can be used as an antioxidant, as a free radicalscavenger, to remove reactive oxygen species (ROS) implicated in manydiseases, making these species unavailable to human tissues. It can alsobe used as an antiinflammatory. A problem is that the currentpreparations are typically large in order to obtain a desired dosage, ormultiple capsules have to be taken, resulting in an inconvenience to theconsumer and, of course, non-compliance. Stable, concentratedformulations, such as solutions, emulsions, microemulsions, andnanoemulsions, would be appreciated by those skilled in the art in orderto increase compliance of administration, as well as thebioavailability, of the components of the extract after administration.

One of skill in the art will appreciate having (i) a sunscreen with avery broad band of absorption with desired strength across the UVA andUVB ranges without requiring the addition of titanium dioxide. Inaddition, the art would appreciate having (ii) a process of producingthe extract without requiring the removal of the extraction solvent,reducing complexity and cost of processing. Moreover, the art would alsoappreciate (iii) an antioxidant dosage form that not only can be useddirectly from the extraction process, but also is potent andconcentrated to a smaller quantity for consumption for increasedcompliance with a variety of administrations and uses. In particular,the art would appreciate having (iv) microemulsion and nanoemulsionformulations that can be easily produced directly from the extractionprocess without requiring further separation of extraction solvents, theemulsions providing an enhancement to the bioavailability of theextracts as well as an increased stability. And, the art would alsoappreciate having (v) extract formulations that can be used incombination as a topically, orally, and/or rectally administeredcomposition for the variety of indications taught herein. Finally, theart will appreciate having extraction methods that can provide all ofthe above while also providing (vi) an extraction process having asignificantly higher yield than state-of-the-art processes.

SUMMARY

The teachings provided herein generally relate to active compositionsthat can be isolated from plants in the Zingiberaceae family for safeuse topically, orally, rectally or vaginally, for example, namely theCurcuma family, and more particularly Curcuma longa L. or “turmeric”. Insome embodiments, the teachings are directed to a composition for use asa sunscreen, the composition comprising an extract of a Curcuma longa L.root.

In some embodiments, the teachings are directed to a pharmaceuticallyacceptable composition comprising a purified liquid extract from theroots of a Curcuma longa L. plant. The liquid extract can comprisecompounds of interest, such as curcumin; demethoxycurcumin;bisdemethoxycurcumin; or, a combination thereof. The liquid extract alsocontains an at least substantially non-toxic extraction solvent thatremoves the compounds of interest from the roots of a Curcuma longa L.plant in an extraction process; and, functions as a pharmaceuticallyacceptable carrier having (i) an acute oral toxicity with an LD50 of atleast 49,700 mg/kg or (ii) an acute dermal toxicity of at least 5000mg/kg; wherein, the extraction solvent was used in the extractionprocess that removed the extract from the roots of the Curcuma longa L.plant and chosen to remain as a component of the pharmaceuticallyacceptable composition.

It should be appreciated that the term “purified” can be used to referto an extract from a Curcuma longa plant, in some embodiments, such thatthe compounds of interest are isolated from the remainder of the plantin a soluble form. As such, one of skill will appreciate that thecompounds of interest can sometimes be accompanied by other componentsof the plant that are carried along with the extract. For example, suchother components can include those selected from the group consisting ofhemicellulose, resins, oils, volatiles, inorganic salts, or anycombination thereof. In some embodiments, the term “purified” can beused to refer to an extract consisting of, or consisting essentially of,any one or any combination of the compounds of interest. In someembodiments, the extract includes the extraction solvent. In someembodiments, an extract “consists essentially of” any one or anycombination of the compounds of interest, where the presence of anyother component from the plant has a negligible effect on the activityof the compounds of interest. The term “negligible effect” can be usedto mean that the activity does not increase or decrease more than about10% when compared to any one or any combination of the compounds ofinterest, respectively, without the other components. In someembodiments, the term “negligible effect” can be used to refer to achange of less that 10%, less than 9%, less than 8%, less than 7%, lessthan 6%, less than 5%, less than 4%, and less than 3%. In someembodiments, the term “negligible effect” can be used to refer to achange ranging from about 3% to about 10%, in increments of 1%. In someembodiments, the other components, for example, the hemicellulose,resins, oils, volatiles, inorganic salts, or any combination thereof,can enhance the activity of the compounds of interest. For example, theactivity of the compounds of interest can be enhanced by an amountranging from about 10% to about 300%, from about 20% to about 200%, fromabout 25% to about 250%, from about 30% to about 300%, from about 35% toabout 275%, from about 40% to about 225%, from about 15% to about 100%,or any range therein in increments of 1%.

It should be appreciated that the term “extract” can be used to refer toa powder form of the compounds of interest, a liquid form of thecompounds of interest, or any one or any combination of the compounds ofinterest in powder or liquid form. One of skill will appreciate that theterm “extract” can be used to refer to the compounds of interest before,during, or after their removal from the plant. In some embodiments, thecompounds of interest can be synthesized chemically using standardmethods known to one of skill, such that they can be synthesized andused alone, or in any combination, by those of skill without use of theextraction methods taught herein.

Any dosage form known to one of skill can be used for administrationsthat include, for example, topical, oral, rectal, or vaginaladministration. In some embodiments, the composition is in a dosage formfor administration topically for any use set-forth herein. In someembodiments, the composition is applied topically as a sunscreen. And,in some embodiments, the composition is in a dosage form foradministration orally. In some embodiments, the dosage form can be acapsule or tablet. The composition can be used as a dietary supplement.In some embodiments, the dietary supplement can function as anantioxidant in a subject.

The teachings are also directed to a pharmaceutically acceptableemulsion comprising a purified liquid extract from the roots of aCurcuma longa L. plant. The emulsion can comprise curcumin;demethoxycurcumin; bisdemethoxycurcumin; or, a combination thereof. Theemulsion includes an at least substantially non-toxic extraction solventthat removes the extract from the roots of a Curcuma longa L. plant inan extraction process; and, functions as a pharmaceutically acceptablecarrier having (i) an acute oral toxicity with an LD50 of at least49,700 mg/kg or (ii) an acute dermal toxicity of at least 5000 mg/kg;wherein, the extraction solvent was (iii) used in the extraction processthat remove the extract from the roots of the Curcuma longa L. plant and(iv) chosen to remain as a component of the pharmaceutically acceptablecomposition. The emulsion comprises a pharmaceutically acceptable oil;and, an emulgent. One of skill will appreciate that an emulsion can beconsidered as a fine dispersion of minute droplets of one liquid inanother in which it is not soluble or miscible. As such, an emulsioncontaining a pharmaceutically acceptable oil and an emulgent can alsocontain water, an alcohol, or any other suitable hydrophilic liquid suchas, for example, polyethylene glycol 200 (PEG200) or polyethylene glycol400 (PEG 400).

The teachings are also directed to methods of preparing thepharmaceutically acceptable compositions taught herein. The methods cancomprise macerating at least a portion of a Curcuma longa L. root for aneffective time in the at least substantially non-toxic extractionsolvent. In these embodiments, the extraction solvent is miscible withphenolic diketones that include curcumin, demethoxycurcumin, andbisdemethoxycurcumin; and, pharmaceutically acceptable as a carrier forthe phenolic diketones. As a carrier, it should have (i) an acute oraltoxicity with an LD50 of at least 49,700 mg/kg or (ii) an acute dermaltoxicity of at least 5000 mg/kg. The macerating includes covering theportion of the root with the extraction solvent. The method alsoincludes separating the extraction solvent from the macerated root tocreate a liquid extract of Curcuma longa L.

The teachings are also directed to a method of preparing thepharmaceutically acceptable emulsions that include the extracts taughtherein. The method comprises macerating at least a portion of a Curcumalonga L. root for an effective time in the at least substantiallynon-toxic extraction solvent that is miscible with phenolic diketonesthat include curcumin, demethoxycurcumin, and bisdemethoxycurcumin; andpharmaceutically acceptable as a carrier for the phenolic diketones, thecarrier having (i) an acute oral toxicity with an LD50 of at least49,700 mg/kg or (ii) an acute dermal toxicity of at least 5000 mg/kg.The method also includes separating the extraction solvent from themacerated root to create a liquid extract of Curcuma longa L. and,emulsifying the liquid extract.

In some embodiments, the emulsifying includes adding a pharmaceuticallyacceptable oil to the liquid extract to create the emulsion of theliquid extract. And, in some embodiments, the emulsifying includesadding an emulgent to the liquid extract to create the emulsion of theliquid extract as described above.

The extraction solvents used in the teachings can be any such extractionsolvent, or any combination of such solvents, known to one of skill thatmeets the criteria. In some embodiments, the extraction solventcomprises polyoxyethylene (20) sorbitan monooleate (TWEEN 80),polyethylene glycol, isopropyl myristate, or a combination thereof. Insome embodiments, the extraction solvent comprises a combination ofTWEEN 80 and isopropyl myristate. In some embodiments, the extractionsolvent comprises a ratio of TWEEN 80:isopropyl myristate ranging fromabout 20:80 to about 50:50.

The pharmaceutically acceptable oil used in the teachings can be anysuch oil, or any combination of oils, known to one of skill that meetsthe criteria. In some embodiments, the pharmaceutically acceptable oilcomprises an oil selected from the group consisting of an animal oil, afish oil, a vegetable oil, or a mineral oil. In some embodiments, thepharmaceutically acceptable oil comprises an edible oil selected fromthe group consisting of olive oil, sunflower oil, sesame oil, almondoil, corn oil, orange oil, lime oil, black pepper oil, nutmeg oil, basiloil, rosemary oil, clove oil, grapefruit oil, fennel oil, coriander oil,bergamot oil, cinnamon oil, lemon oil, peppermint oil, garlic oil, thymeoil, marjoram oil, lemongrass oil, ginger oil, cardamom oil, liquidparaffin, cotton seed oil, peanut oil, nut oil, soy oil, rapeseed oil,vitamin E oil, Vitamin E TPGS oil, fish oil, tallow-derived oil,silicone oil, castor oil, squalene oil, or any mixture thereof.

The teachings are also directed to kits that contain a combination oftopical, oral, or rectal dosage forms for administrations to a subject.In some embodiments, the kit is for protecting a dermal tissue from UVAand UVB exposure, the kit comprising an extract of a Curcuma longa L.root in an oral dosage form; an extract of a Curcuma longa L. root in atopical dosage form; and, instructions for administration of the topicaldosage form, the oral dosage form, or a combination of the topical andoral dosage forms.

The extracts taught herein can be used for a variety of treatments. Insome embodiments, the teachings are directed to a method of treating askin to prevent or inhibit exposure of the skin to UVA and UVB, themethod comprising topically administering an effective amount of acomposition taught herein to a dermal tissue of a subject. In someembodiments, the methods further comprise orally administering aneffective amount of an oral dosage form of a composition taught hereinto a subject to systemically treat a disease or disorder, such as adisease or disorder taught herein. In some embodiments, the methodsfurther comprise orally administering an effective amount of an oraldosage form of a composition taught herein to a subject as a dietarysupplement. In some embodiments, the methods further comprise orallyadministering an effective amount of an oral dosage form of acomposition taught herein to a subject in combination with the topicaladministration.

In some embodiments, the teachings are directed to a method of treatingan inflammation of a tissue of subject, the method comprisingadministering an effective amount of a composition taught herein to atissue of the subject. In some embodiments, the teachings are directedto treating a wounded tissue, the method comprising administering aneffective amount of a composition taught herein to a tissue of thesubject.

Any tissue that can make contact with one or more active components ofan extract taught herein can be treated. The tissue can be, for example,connective, muscle, nervous, and/or epithelial tissue. In someembodiments, the tissue is a dermal tissue. In some embodiments, thetissue is a mucosal tissue. And, in some embodiments, the tissue isgastrointestinal tissue. In some embodiments, a first tissue makescontact with one or more active components of an extract taught hereinand a second tissue realizes a benefit as a secondary effect.

One of skill reading the teachings that follow will appreciate that theconcepts can extend into additional embodiments that go well-beyond aliteral reading of the claims, the inventions recited by the claims, andthe terms recited in the claims.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A-1F compare the absorption spectra of representative currentstate-of-the-art sunscreen agents to the absorption spectra of Curcumalonga L., according to some embodiments.

FIGS. 2A and 2B compare the antioxidant activity of the TWEEN extract ofCurcumin longa to the antioxidant activity of resveratrol, according tosome embodiments.

DETAILED DESCRIPTION

The teachings provided herein generally relate to active extracts,compositions that can be isolated from plants in the Zingiberaceaefamily for use topically, orally, rectally, or vaginally, for example,namely the Curcuma genus, and more particularly Curcuma longa L. or“turmeric”. Any active extract obtained from the Zingiberaceae familycan be used in aspects of the teachings herein. In some embodiments, theteachings are directed to a composition for use as a sunscreen, thecomposition comprising an extract of a Curcuma longa L. root.

In some embodiments, an extract from the Curcuma, or Curcuma L., genuscan be used. In some embodiments, the extract can be from Curcumaaeruginosa, Curcuma albicoma, Curcuma albiflora, Curcuma alismatifolia,Curcuma amada Roxb, Curcuma amarissima, Curcuma angustifolia, Curcumaaromatica, Curcuma attenuata, Curcuma aurantiaca, Curcuma australasica,Curcuma bakeriana, Curcuma bicolor, Curcuma bhatii, Curcuma burttii,Curcuma caesia, Curcuma ceratotheca, Curcuma chuanezhu, Curcumachuanhuangjiang, Curcuma chuanyujin, Curcuma cochinchinensis, Curcumacodonantha, Curcuma coerulea, Curcuma colorata, Curcuma comosa, Curcumacoriacea, Curcuma decipiens, Curcuma domestica, Curcuma ecalcarata,Curcuma euchroma, Curcuma ecomata, Curcuma elata, Curcuma exigua,Curcuma ferruginea, Curcuma flaviflora, Curcuma glans, Curcumagracillima, Curcuma grandiflora, Curcuma haritha, Curcuma harmandii,Curcuma heyneana, Curcuma inodora, Curcuma inodora, Curcumakarnatakensis, Curcuma kudagensis Velayudhan, Curcuma kwangsiensis,Curcuma lanceolata, Curcuma larsenii, Curcuma latiflora, Curcumalatifolia, Curcuma leucorrhiza, Curcuma loerzingii, Curcuma longa L,Curcuma longa, Curcuma longispica, Curcuma malabarica, Curcumameraukensis, Curcuma mutabilis, Curcuma neilgherrensis, Curcumanilamburensis, Curcuma oligantha, Curcuma ornata, Curcuma parviflora,Curcuma parvula, Curcuma peethapushpa, Curcuma petiolata, Curcumaphaeocaulis, Curcuma pierreana, Curcuma plicata, Curcuma porphyrotaenia,Curcuma prakasha, Curcuma pseudomontana, Curcuma purpurascens, Curcumapurpurea, Curcuma raktakanta, Curcuma reclinata, Curcuma rhabdota,Curcuma rhomba, Curcuma roscoeana, Curcuma rubescens, Curcumarubrobracteata, Curcuma sattayasaii, Curcuma sichuanensis, Curcumasingularis, Curcuma sparganiifolia, Curcuma stenochila, Curcumastrobilifera, Curcuma sulcata, Curcuma sumatrana, Curcuma sylvatica,Curcuma thalakaveriensis, Curcuma thorelii, Curcuma trichosantha,Curcuma vamana, Curcuma vellanikkarensis, Curcuma wenyujin, Curcumawenchowensis, Curcuma xanthorrhiza, Curcuma yunnanensis, Curcumazedoaria, Curcuma zedoaroides, and any combination thereof.

In some embodiments, the extract can be from Curcuma aeruginosa, Curcumadomestica, Curcuma longa, Curcuma manga, Curcuma xanthorriza, or acombination thereof. In some embodiments, the extract can be from Alpinagalaiga, Amonum kepulaga, Phoeomera speciosa, Zinger cassumunar, or acombination thereof.

The compositions provided herein can be referred to as extracts,compositions, compounds, agents, active agents, bioactive agents,supplements, drugs, and the like. In some embodiments, the terms“composition,” “compound,” “agent,” “active”, “active agent”, “bioactiveagent,” “supplement,” and “drug” can be used interchangeably and, itshould be appreciated that, a “formulation” can comprise any one or anycombination of these. Likewise, in some embodiments, the composition canalso be in a liquid or dry form, where a dry form can be a powder formin some embodiments, and a liquid form can include an aqueous ornon-aqueous component. Moreover, the term “bioactivity” can refer to thefunction of the compound when administered orally, topically, orrectally to a subject.

In some embodiments, the term “target site” can be used to refer to aselect location on or in a subject that could benefit from anadministration of a compound taught herein, either topically or orally.In some embodiments, a target can include any site of action in whichthe agent's activity, such as antioxidant activity, anti-inflammatoryactivity, or UVA and UVB blocking activity, can serve a benefit to thesubject. The target site can be a healthy or damaged tissue of asubject. As such, the teachings include a method of administering one ormore compounds taught herein to a healthy or damaged tissue, dermal,mucosal, gastrointestinal or otherwise.

One of skill will appreciate that the compositions or formulationsshould remain stable, or at least substantially stable, until useful oractivated, and this can relate to a shelf life, or a time betweencreation and administration of the composition, or some combinationthereof. In some embodiments, the composition is stable, orsubstantially stable, when usable as intended within a reasonable amountof time, a time that is considered reasonable by one of skill for theapplications taught herein. In some embodiments, the composition shouldbe usable within a reasonable time from the making to the administrationof the composition and, in some embodiments, the composition should havea reasonable commercial shelf life, a shelf life that is consideredreasonable to one of skill. A reasonable shelf life can be at least 6months, at least 1 year, at least 18 months, at least 2 years, at least3 years, or any time in-between in increments of about 1 month, in someembodiments.

In some embodiments, a composition or formulation can be considered as“stable” if it loses less than 10% of its original activity. In someembodiments, a composition or formulation can be considered as stable ifit loses less than 5%, 3%, 2%, or 1% of its original activity. In someembodiments, a composition or formulation can be considered as“substantially stable” if it loses greater than about 10% of itsoriginal activity, as long as the composition can perform it's intendeduse to a reasonable degree of efficacy. In some embodiments, thecomposition can be considered as substantially stable if it losesactivity at an amount greater than about 12%, about 15%, about 25%,about 35%, about 45%, about 50%, about 60%, or even about 70%. Theactivity loss can be measured by comparing activity at the time ofpackaging to the activity at the time of administration, and this caninclude a reasonable shelf life. In some embodiments, the composition isstable or substantially stable, if it remains useful for a periodranging from 3 months to 3 years, 6 months to 2 years, 1 year, or anytime period therein in increments of about 1 month.

Methods of Use

The extracts taught herein can be used for a variety of treatments,preventative, ameliorative, or otherwise, as well as for use as adietary supplement. The uses can include medicinal purposes, as a healthsupplement, a nutritional composition, a prophylactic, or a treatment ofan existing condition. In some embodiments, any tissue that can makecontact with one or more active components of an extract taught hereincan be treated. In some embodiments, a tissue can have a desirablesecondary effect from one or more of the active components of an extracttaught herein making contact elsewhere in the subject, such that one ormore of the active components can contact a first tissue, whereas asecond tissue realizes a beneficial effect. For example, the firsttissue can be a stomach lining, and the second tissue can realize thedesirable effect of a release of a neurotransmitter or a neuroimpulse.The tissue can be, for example, connective, muscle, nervous, and/orepithelial tissue. In some embodiments, the tissue is a dermal tissue.In some embodiments, the tissue is a mucosal tissue. And, in someembodiments, the tissue is gastrointestinal tissue.

The terms “treat,” “treating,” and “treatment” can be usedinterchangeably in some embodiments and refer to the administering orapplication of the compositions and formulations taught herein,including such administration as a health or nutritional supplement, andall administrations directed to the prevention, inhibition, ameliorationof the symptoms, or even a cure of a condition taught herein. The terms“disease,” “condition,” “disorder,” and “ailment” can be usedinterchangeably in some embodiments. The term “subject” and “patient”can be used interchangeably in some embodiments and refer to an animalsuch as a mammal including, but not limited to, non-primates such as,for example, a cow, pig, horse, cat, dog, rat and mouse; and primatessuch as, for example, a monkey or a human. As such, the terms “subject”and “patient” can also be applied to non-human biologic applicationsincluding, but not limited to, veterinary, companion animals, commerciallivestock, and the like.

In some embodiments, the teachings are directed to a method of treatinga skin to prevent or inhibit exposure of the skin to UVA and UVB, themethod comprising topically administering an effective amount of acomposition taught herein to a dermal tissue of a subject. In someembodiments, the methods further comprise orally administering aneffective amount of an oral dosage form of a composition taught hereinto a subject to systemically treat a disease or disorder, such as adisease or disorder taught herein. In some embodiments, the methodsfurther comprise orally administering an effective amount of an oraldosage form of a composition taught herein to a subject as a dietarysupplement. In some embodiments, the methods further comprise orallyadministering an effective amount of an oral dosage form of acomposition taught herein to a subject in combination with the topicaladministration. In some embodiments, the teachings are directed to amethod of treating an inflammation of a tissue of subject, the methodcomprising administering an effective amount of a composition taughtherein to a tissue of the subject. In some embodiments, the teachingsare directed to treating a wounded tissue, the method comprisingadministering an effective amount of a composition taught herein to atissue of the subject.

An “effective amount” of a compound can be used to describe atherapeutically effective amount or a prophylactically effective amount.An effective amount can also be an amount that ameliorates the symptomsof a disease. A “therapeutically effective amount” can refer to anamount that is effective at the dosages and periods of time necessary toachieve a desired therapeutic result and may also refer to an amount ofactive compound, prodrug or pharmaceutical agent that elicits anybiological or medicinal response in a tissue, system, or subject that issought by a researcher, veterinarian, medical doctor or other clinicianthat may be part of a treatment plan leading to a desired effect. Insome embodiments, the therapeutically effective amount should beadministered in an amount sufficient to result in amelioration of one ormore symptoms of a disorder, prevention of the advancement of adisorder, or regression of a disorder. In some embodiments, for example,a therapeutically effective amount can refer to the amount of an agentthat provides a measurable response of at least 5%, at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 55%, at least 60%, atleast 65%, at least 70%, at least 75%, at least 80%, at least 85%, atleast 90%, at least 95%, or at least 100% of a desired action of thecomposition.

In cases of the prevention or inhibition of the onset of a disease ordisorder, or where an administration is considered prophylactic, aprophylactically effective amount of a composition or formulation taughtherein can be used. A “prophylactically effective amount” can refer toan amount that is effective at the dosages and periods of time necessaryto achieve a desired prophylactic result, such as prevent the onset of asunburn, an inflammation, allergy, nausea, diarrhea, infection, and thelike. Typically, a prophylactic dose is used in a subject prior to theonset of a disease, or at an early stage of the onset of a disease, toprevent or inhibit onset of the disease or symptoms of the disease. Aprophylactically effective amount may be less than, greater than, orequal to a therapeutically effective amount.

In some embodiments, a therapeutically or prophylactically effectiveamount of a composition may range in concentration from about 0.01 nM toabout 0.10 M; from about 0.01 nM to about 0.5 M; from about 0.1 nM toabout 150 nM; from about 0.1 nM to about 500 μM; from about 0.1 nM toabout 1000 nM, 0.001 μM to about 0.10 M; from about 0.001 μM to about0.5 M; from about 0.01 μM to about 150 μM; from about 0.01 μM to about500 μM; from about 0.01 μM to about 1000 nM, or any range therein. Insome embodiments, the compositions may be administered in an amountranging from about 0.005 mg/kg to about 100 mg/kg; from about 0.005mg/kg to about 400 mg/kg; from about 0.01 mg/kg to about 300 mg/kg; fromabout 0.01 mg/kg to about 250 mg/kg; from about 0.1 mg/kg to about 200mg/kg; from about 0.2 mg/kg to about 150 mg/kg; from about 0.4 mg/kg toabout 120 mg/kg; from about 0.15 mg/kg to about 100 mg/kg, from about0.15 mg/kg to about 50 mg/kg, from about 0.5 mg/kg to about 10 mg/kg, orany range therein, wherein a human subject is often assumed to averageabout 70 kg. In addition to use as a sunscreen as provided herein, theextracts taught herein can provide a number of therapeutic benefits.Such benefits can include, for example, free radical scavenger andantioxidant, inhibiting lipid peroxidation and oxidative DNA damage;anti-inflammatory activity; neurological treatments for Alzheimer'sdisease (anti-amyloid and other effects), Parkinson's disease, and otherneurological disorders; anti-arthritic treatment; anti-ischemictreatment; treatments for multiple myeloma and myelodysplasticsyndromes; psoriasis treatments (topically and orally); cystic fibrosistreatments; treatments for liver injury and alcohol-induced liverdisease; multiple sclerosis treatments; antiviral treatments, includinghuman immunodeficiency virus (HIV) therapy; treatments of diabetes;cancer treatments; and, reducing risk of heart disease; to name a few.

As such, the use of Curcuma longa L. extract as a sunscreen agent hasthe potential of providing additional therapeutic benefits that couldfar exceed merely protecting the skin from UV the rays of the sun. Onthe skin, this effect may aid in preventing reactions in individualswith sensitive skin. The limitation of these reactions may help influshing, blushing, rosacea and similar disorders in such individuals.Given the above, the application of the sunscreen may provide beneficialeffects to sportsmen who apply it since minor scratches and injuriessustained during a sports game may heal faster. It would have a similaradvantage to gardeners, those at the beach, hikers and so on. Theextracts can be used, for example, as a dietary supplement or apharmaceutically acceptable dosage form.

In some embodiments, the extracts taught herein can be used as anantioxidant by the oral route, as a free radical scavenger, to removereactive oxygen species (ROS) implicated in many diseases, making thesespecies unavailable to human tissues. In some embodiments, the dietarysupplement can function as an antioxidant in a subject. The compositioncan be used as a dietary supplement in some embodiments.

The compositions or formulations are also useful in treatinginflammations. In some embodiments, they can be administered fortreating inflammations of any tissue such as, for example, skin,mucosal, or gastrointestinal inflammations in a subject, in which theycan be administered, for example, topically, orally, rectally, orvaginally to prevent, treat, inhibit, or ameliorate the symptoms of aninflammation of the tissue.

In some embodiments, the compositions and formulations can be used toprevent, treat, ameliorate the symptoms of, or even cure, an acute orchronic gastrointestinal condition. Such conditions can include, but arenot limited to, hyperacidity, colitis, irritable bowel syndrome, Crohn'sdisease, necrotic enteritis, functional colonic diseases, malabsorption,a peptic ulcer, gastro-esophageal reflux disease, ulcerative colitis,and diverticulitis. In some embodiments, the compositions andformulations can be used to reduce mucosal tissue inflammation,dysfunction, or damage. Such conditions can be induced, for example, bydrug side effects, chemotherapy, dysbiosis, radiation, changes in normalflora, hyperimmunity, autoimmune reactions, immune deficiencies,nervousness, allergies, chemical irritation, and stress. In someembodiments, the symptoms of a gastrointestinal condition can include,for example, diarrhea, dehydration, malnutrition, constipation, nausea,and/or cramping. And, in some embodiments, the symptoms of agastrointestinal condition can be temporary and include acid irritation,indigestion, bloating, cramps, spasmodic peristalsis, diarrhea, andconstipation. Administering the compositions or formulations for thetreatment and/or management of gastrointestinal conditions can beconsidered a nutritional or health supplement, in some embodiments.

The compositions or formulations are also useful in treating wounds.Generally speaking, they can be administered to protect, promotehealing, or improve function of skin or mucosa. In some embodiments, forexample, a wound and a chronic inflammatory condition can be treatedincluding, but not limit to, a wound by (i) physical damage, (ii) adiabetic skin lesion, (iii) a bed sore, (iv) a burn, (v) a cold sore,(vi) psoriasis, (vii) eczema, and (viii) dermatological inflammationcaused by pathogens, to name a few.

Making and Administering the Extract

Making the Curcuma longa Extract

Generally speaking, at least a portion of a root of Curcuma longa L.,for example, can be crushed and then soaked in a solvent to create amacerate. The macerate can be filtered and the residue can bere-extracted multiple times. The extract can be adjusted for potency andstored in amber bottles. The extract may be prepared as an alcoholicextract, an oily extract or another type of organic solvent may be used.The extract may be used as an alcoholic solution or the solvent may beevaporated and the dry powder incorporated into suitable formulationsfor human use. The extraction may also be done using oils suitable forhuman use as the solvent with the advantage of incorporating the extractdirectly into sunscreen cream or lotion formulations.

In some embodiments, the solvent can be a lower alkanol. Theconcentrations of lower alkanol used in the extractions can range from,for example, 20%-30%, 30%-40%, 40%-50%, 50%-60%, 60%-70%, 70%-80%,80%-90%, or any concentration therein. The alkanol can be used inadditional washes for further purification of the extract. An extractprecipitate, for example, can be washed with more of the lower alkanol,where a typical wash may include, for example, three washes with 95%ethanol. The precipitate can then be suspended in water at aconcentration suitable for further processing such as, for example,about 18-20% weight/volume. Re-precipitation by again adding a loweralkanol in the water can be used to remove additional materials that arenot water-soluble. The supernatant can be precipitated with a higherconcentration of the lower alkanol.

In some embodiments, the higher concentration of the lower alkanol canbe, for example, ethanol. Ethanol appears to be a favorable extractionsolvent, at least with regard to separating the extract from the plant,as a comparatively high yield is obtained when compared to otherstate-of-the-art methods. In addition, the extract is not viscous whichmakes filtration easier. Since ethanol is a suitable extraction solventand oils can be incorporated into formulations, it is also possible tore-extract an ethanol extract into suitable oils, such as sesame seedoil. For the latter, several extraction steps may be necessary.

In some embodiments, 40-80% ethanol, 60-70% ethanol, or 50-60% ethanolcan be used to create a crude extract. In some embodiments, the ethanolcan be added at a concentration of about 70% ethanol at about roomtemperature, and the extraction can be done in steps by first using alower concentration of about 35% ethanol in a first step, and then usinga higher concentration of about 70% ethanol in a second step.

In some embodiments, the extract can be dried. The application of heat,however, should be carefully performed, as the extracts degrade whensubject to heat and, thus, the activity of the extract can bedeleteriously affected. In general, it is better to heat for theshortest time possible to limit degradation. In some embodiments, theextract can be dried for a time ranging from about 30 seconds to about 1minute, from about 1 minute to about 5 minutes, from about 5 minute toabout 10 minutes, from about 10 minute to about 20 minutes, from about20 minute to about 30 minutes, from about 30 minute to about 40 minutes.In some embodiments, the extract can be dried for a time ranging for atime ranging from about 30 minutes to 1 hour or more, depending on theconditions used. In some embodiments, heat can be applied attemperatures below about 90° C., ranging from about 30° C. to about 80°C., from about 40° C. to about 70° C., from about 50° C. to about 60°C., from about 70° C. to about 80° C., or any range therein. In someembodiments, the heating can be at a temperature ranging from 25° C. to70° C. in any increment of about 5° C. It should be appreciated that anyprocess known to one of skill that avoids any heating, or at leastexcessive heating can be used, including, for example, drying in adessicator at room temperature, spray drying, vacuum drying,freeze-drying, critical point drying, solvent exchange, and anycombination thereof known to one of skill.

In some embodiments, the extract can be filtered. For example, anextract can be re-dissolved, and brought to a suitable concentration forfiltration to further remove unwanted materials. In some embodiments,the suitable concentration for filtration can be about 1%, about 2%,about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%,about 10%, about 12%, about 14% or any concentration therein inincrements of 0.2%.

In some embodiments, the extraction is done using an oil or othersolvent. In some embodiments, the solvent can include a surfactant.Example surfactants include anionic, cationic, zwitterionic, andnonionic surfactants. An example of cationic surfactants includes sodiumdodecylsulfate (SDS). An example of anionic surfactants includescetylpyridinium bromide. An example of zwitterionic surfactants includesdipalmitoylphosphatidylcholine (lecithin). An example of noninionicsurfactants includes polyoxyethylene(4) lauryl ether (BRIJ 30). In someembodiments, SPAN- or TWEEN-type surfactants can be used, including SPAN60, SPAN 80, TWEEN 60, and TWEEN 80. In fact, any one or any combinationof the numerous surfactants known to one of skill can be used to theextent that there is reason to believe that such use would be consistentwith the teachings provided herein.

In some embodiments, the solvent can include an oil. The oil and/orsurfactant can be used to extract the root, Curcuma longa L., bycrushing the root and soaking the crushed root in the solvent to form amacerate. The macerate is then filtered to leave an extract. Themacerate will often contain more extract and, as such, the residue maybe re-extracted multiple times in some embodiments, and any one orcombination of solvents can be used to obtain the extract from the root.In some embodiments, sesame seed oil can be used, as it is considered atleast substantially non-toxic and light, such that the extract can befiltered easily and not display excessive oiliness when applied to theskin.

Method of preparing the pharmaceutically acceptable compositions taughtherein can comprise macerating at least a portion of a Curcuma longa L.root for an effective time in the at least substantially non-toxicextraction solvent. In these embodiments, the extraction solvent ismiscible with phenolic diketones that include curcumin,demethoxycurcumin, and bisdemethoxycurcumin; and, pharmaceuticallyacceptable as a carrier for the phenolic diketones. The method alsoincludes separating the extraction solvent from the macerated root tocreate a liquid extract of Curcuma longa L.

The teachings are also directed to a method of preparing thepharmaceutically acceptable emulsions that include the extracts taughtherein. In some embodiments, the emulsifying includes adding apharmaceutically acceptable oil to the liquid extract create theemulsion of the liquid extract. And, in some embodiments, theemulsifying includes adding an emulgent to the liquid extract create theemulsion of the liquid extract. As such, the method can comprisemacerating at least a portion of a Curcuma longa L. root for aneffective time in the at least substantially non-toxic extractionsolvent that is miscible with phenolic diketones that include curcumin,demethoxycurcumin, and bisdemethoxycurcumin; and pharmaceuticallyacceptable as a carrier for the phenolic diketones. The method alsoincludes separating the extraction solvent from the macerated root tocreate a liquid extract of Curcuma longa L. and, emulsifying the liquidextract.

The term “at least substantially non-toxic” can be used to mean that theextraction solvent is substantially less toxic than ethanol, such thatone of skill would be able to use at least 50%, 75%, 100%, 200%, or 300%more of the solvent as compared to ethanol in a formulation for use as atopical, oral, or rectal formulation. The guidelines for use can followthe US FDA regulations. In some embodiments, the extraction solvent has(i) an acute oral toxicity with an LD50 of at least 49,700 mg/kg or (ii)an acute dermal toxicity of at least 5000 mg/kg.

The extraction solvents used in the teachings can be any such extractionsolvent, or any combination of such solvents, known to one of skill thatmeets the criteria. In some embodiments, the extraction solventcomprises polyoxyethylene (20) sorbitan monooleate (TWEEN 80),polyethylene glycol, isopropyl myristate, or a combination thereof. Insome embodiments, the extraction solvent comprises a combination ofTWEEN 80 and isopropyl myristate. In some embodiments, the extractionsolvent comprises a ratio of TWEEN 80:isopropyl myristate ranging fromabout 20:80 to about 50:50.

The pharmaceutically acceptable oil used in the teachings can be anysuch oil, or any combination of oils, known to one of skill that meetsthe criteria. In some embodiments, the pharmaceutically acceptable oilcomprises an oil selected from the group consisting of an animal oil, afish oil, a vegetable oil, or a mineral oil. In some embodiments, thepharmaceutically acceptable oil comprises an edible oil selected fromthe group consisting of olive oil, sunflower oil, sesame oil, almondoil, corn oil, orange oil, lime oil, black pepper oil, nutmeg oil, basiloil, rosemary oil, clove oil, grapefruit oil, fennel oil, coriander oil,bergamot oil, cinnamon oil, lemon oil, peppermint oil, garlic oil, thymeoil, marjoram oil, lemongrass oil, ginger oil, cardamom oil, liquidparaffin, cotton seed oil, peanut oil, nut oil, soy, rapeseed oil,vitamin E oil and derivatives thereof, including Vitamin E TPGS (d-alphatocopheryl polyethylene glycol 1000 succinate), fish oil, tallow-derivedoil, silicone oil, castor oil, squalene oil, or any mixture thereof.

Formulations

Curcuma longa L. can be applied to a target tissue non-parenterally. Insome embodiments, the compositions or formulations can be applied atleast topically, orally, rectally, or vaginally, for example. And, anysuitable dosage form known to one of skill can be used for topical,oral, rectal, or vaginal administration.

The design of the formulations can include, for example, (i) identifyingthe condition or use; (ii) identifying the target site; and (ii)matching a dosage form for administration to the target site.Identifying the target site includes, for example, select a targettissue for treatment, such as a healthy or damaged tissue at which theformulation can be applied to prevent, inhibit, otherwise treat, orameliorate the symptoms of a condition. Once the use, the target site,and the method of administration have been chosen, one of skill canreadily select a dose, which will vary according to any of a variety offactors known the person of skill including, but not limited to,environmental conditions present at the site of use, for example,sunlight, heat, water, pH, gastric acids, and the like. In someembodiments, this formulation can be administered for uses in animalsthat are non-humans.

In some embodiments, the composition is in a dosage form foradministration topically for any use set-forth herein. Topicaladministration can be used on any tissue that can benefit from a topicalapplication, including the skin, mucous membranes, and gastrointestinaltract. For topical administration, suitable formulations may include abiocompatible oil, wax, gel, powder, emulsion, polymer, or other liquidor solid carriers. Such formulations may be administered by applyingdirectly to affected tissues. For example, a cream formulation can beadministered to a target area on the skin. Transdermal administrationsare also possible, including percutaneous absorption of the Curcumacompounds through the skin. Transdermal formulations can includepatches, ointments, creams, gels, salves, and the like. In someembodiments, the composition is applied topically as a sunscreen.

In some embodiments, the composition is in a dosage form foradministration orally. In some embodiments, the dosage form can be acapsule or tablet. Powder forms can also be produced from the liquidextract with the methods taught herein, and the powder can offer evengreater ease of consumption when manufactured into dosage forms known toone of skill, such as filled into capsules or compressed into tablets.Powders can also offer greater stability over liquids in someembodiments.

In some embodiments, the extract can be an oil extract or a surfactantextract that may be filled into capsules, or it can be an alcoholextract that may be dried to a powder, and the powder filled intocapsules or made into tablets. In some embodiments, the extract can beextracted with an edible oil as an extraction solvent, as the oilextract can be emulsified and is at least substantially non-toxic. Sincethe extractable components of the root are highly oil soluble, theyremain predominantly in the oil phase of the emulsion which aids thestability of the product. The liquid extracts that are filled intocapsules are sealed with a gelatin or hydroxypropylmethylcellulose(HPMC) solution, depending on whether the capsule shell is comprised ofgelatin or hydroxypropylmethylcellulose. The seal substantially preventsleakage of the liquid from the shell.

In some embodiments, the compositions or formulations can beadministered in a sustained release formulation, and the formulation caninclude one or more agents in addition to the compositions taughtherein. In some embodiments, the sustained release formulations canreduce the dosage and/or frequency of the administrations of such agentsto a subject.

In some embodiments, the compositions or formulations can beadministered in conjunction with at least one other therapeutic agentfor the condition being treated. The amounts of the agents can bereduced, even substantially, such that the amount of the agent or agentsdesired is reduced to the extent that a significant response is observedfrom the subject. A “significant response” can include, but is notlimited to, a reduction or elimination of a symptom, a visible increasein a desirable therapeutic effect, a faster response to the treatment, amore selective response to the treatment, or a combination thereof. Insome embodiments, the other therapeutic agent can be administered, forexample, in an amount ranging from about 0.1 μg/kg to about 1 mg/kg,from about 0.5 μg/kg to about 500 μg/kg, from about 1 μg/kg to about 250μg/kg, from about 1 μg/kg to about 100 μg/kg from about 1 μg/kg to about50 μg/kg, or any range therein. Combination therapies can beadministered, for example, for 30 minutes, 1 hour, 2 hours, 4 hours, 8hours, 12 hours, 18 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6days, 7 days, 8 days, 9 days, 10 days, 2 weeks, 3 weeks, 4 weeks, 6weeks, 3 months, 6 months 1 year, any combination thereof, or any amountof time considered desirable by one of skill. The agents can beadministered concomitantly, sequentially, or cyclically to a subject.Cycling therapy involves the administering a first agent for apredetermined period of time, administering a second agent or therapyfor a second predetermined period of time, and repeating this cyclingfor any desired purpose such as, for example, to enhance the efficacy ofthe treatment. The agents can also be administered concurrently. Theterm “concurrently” is not limited to the administration of agents atexactly the same time, but rather means that the agents can beadministered in a sequence and time interval such that the agents canwork together to provide additional benefit. Each agent can beadministered separately or together in any appropriate form using anyappropriate means of administering the agent or agents. One of skill canreadily select the frequency, duration, and perhaps cycling of eachconcurrent administration.

Having the Curcumin compounds in the form of extracts, dry or liquid,allows for ease of the selection of dose and concentration in dosageforms. A range of other agents or additives may be included to provideadditional control over a desired therapeutic effect, cosmetic appeal,or perhaps aid in compliance for a recommended use. Stable, concentratedformulations, such as solutions, emulsions, microemulsions, andnanoemulsions would, for example, potentially increase compliance ofadministration and bioavailability of the components of the extractafter administration. A desirable therapeutic effect can include, forexample, increasing bioavailability of the extract through topical useor consumption, providing additional therapeutic effect through additionof a second agent, adjusting the sun protection factor of a sunscreen(spf), and the like. In a sunscreen, for example, the emulsion can beformulated as small globules to provide more complete coverage of theskin. In some embodiments, the globule size can be used to adjust UVprotection and improve the stability of an emulsion. A cosmetic appealcan include, for example, providing the extract as a cream or lotion,adjusting the oiliness of a composition, adding a pleasing scent ormoisturizer, and the like. Aiding in compliance can include, forexample, adjusting the dosage to be in a form that facilitatescompliance of patients by offering a small dosage form for consumptionby concentrating the extract or, perhaps, offering a time-delay dosageform to reduce the frequency of patient intake that is desired toachieve a particular therapeutic effect.

In some embodiments, a pharmaceutically acceptable composition willcomprise a purified and/or isolated liquid extract from the roots of aCurcuma longa L. plant. The liquid extract can comprise curcumin;demethoxycurcumin; bisdemethoxycurcumin; or, a combination thereof. Theliquid extract also contains an at least substantially non-toxicextraction solvent that removes the extract from the roots of a Curcumalonga L. plant in an extraction process; and, functions as apharmaceutically acceptable carrier; wherein, the extraction solvent wasused in the extraction process that removed the extract from the rootsof the Curcuma longa L. plant and chosen to remain as a component of thepharmaceutically acceptable composition.

In some embodiments, an oily extract can be emulsified using, forexample, a surfactant as an emulgent. As such, a pharmaceuticallyacceptable emulsion comprising a purified liquid extract from the rootsof a Curcuma longa L. plant can be made. The emulsion can comprisecurcumin; demethoxycurcumin; bisdemethoxycurcumin; or, a combinationthereof. The emulsion includes an at least substantially non-toxicextraction solvent that removes the extract from the roots of a Curcumalonga L. plant in an extraction process; and, functions as apharmaceutically acceptable carrier having (i) an acute oral toxicitywith an LD50 of at least 49,700 mg/kg or (ii) an acute dermal toxicityof at least 5000 mg/kg; wherein, the extraction solvent was (i) used inthe extraction process that remove the extract from the roots of theCurcuma longa L. plant and (ii) chosen to remain as a component of thepharmaceutically acceptable composition. The emulsion comprises apharmaceutically acceptable oil; and, an emulgent.

Example surfactants include anionic, cationic, zwitterionic, andnonionic surfactants. An example of cationic surfactants includes sodiumdodecylsulfate (SDS). An example of anionic surfactants includescetylpyridinium bromide. An example of zwitterionic surfactants includesdipalmitoylphosphatidylcholine (lecithin). An example of noninionicsurfactants includes polyoxyethylene(4) lauryl ether (BRIJ 30). In someembodiments, SPAN- or TWEEN-type surfactants can be used, including SPAN60, SPAN 80, TWEEN 60, and TWEEN 80. In fact, any one or any combinationof the numerous surfactants known to one of skill can be used to theextent that there is reason to believe that such use would be consistentwith the teachings provided herein.

In some embodiments, the pH of the formulations can be adjusted. The pHof the formulations are an improvement over the art, as the pH can beadjusted. In some embodiments, the pH can be adjusted to that of theskin, for example. Since the pH of skin is approximately 4.2 to 5.5,depending on the individual and area of the body, the pH of a topicalformulation can range from about 4 to about 6.5 in some embodiments,from about 4.3 to about 5.8 in some embodiments, from about 4.5 to about5.5 in some embodiments, or any range therein. In some embodiments, thepH can be 3.8, 4.0, 4.2, 4.4, 4.6, 4.8, 5.0, 5.2, 5.4, 5.6, 5.8, 6.0,6.2, 6.4, 6.6, or any increment of about 0.1 therein.

In some embodiments, the viscosity of the formulations can be adjusted.The viscosity of the formulations is an improvement over the art, as theviscosity can be adjusted. For example, the viscosity can be adjusted bychoice of solvent or oil, for example, or temperature used in processingas, generally, temperature effect on viscosity is temporary. In someembodiments, the viscosity can be adjusted by warming a desired oil, aslong as the heat is not so high as to cause degradation of the Curcumalonga extract. In some embodiments, an oil can be heated up to about 65°C., about 55° C., about 45° C., about 35° C., or any temperature thereinin increments of about 1° C.

Storage of the extract can be in dry form or liquid form. In liquidform, the extract can be adjusted for concentration to vary potency.And, whether dry or liquid form, the extracts can be stored in amberbottles for stability.

Administration

Any administration vehicle known to one of skill to be suitable foradministration of the compounds, compositions, and formulations taughtherein can be used. A “vehicle” can refer to, for example, a diluent,excipient or carrier with which a compound is administered to a subject.

The terms “administration” or “administering” can be used to refer to amethod of incorporating a composition into or onto the cells or tissuesof a subject, either in vivo or ex vivo to test the activity of asystem, as well as to diagnose, prevent, treat, or ameliorate a symptomof a disease or condition. In one example, a compound can beadministered to a subject in vivo using any means of administrationtaught herein. In another example, a compound can be administered exvivo by combining the compound with cell tissue from the subject forpurposes that include, but are not limited to, assays for determiningutility and efficacy of a composition. And, of course, the compositionscan be used in vitro to test their stability, activity, toxicity,efficacy, and the like. When the compound is incorporated in the subjectin combination with one or active agents, the terms “administration” or“administering” can include sequential or concurrent incorporation ofthe compound with the other agents such as, for example, any agentdescribed above. A composition can be formulated, in some embodiments,to be compatible merely with its intended route of administration.

The compounds can be administered in dosage units. The term “dosageunit” can refer to discrete, predetermined quantities of a compound thatcan be administered as unitary dosages to a subject. A predeterminedquantity of active compound can be selected to produce a desiredtherapeutic effect and can be administered with a pharmaceuticallyacceptable carrier. The predetermined quantity in each unit dosage candepend on factors that include, but are not limited to, (a) the uniquecharacteristics of the active compound and the particular therapeuticeffect to be achieved, and (b) the limitations inherent in the art ofcreating and administering such dosage units.

A “pharmaceutically acceptable carrier” is a diluent, adjuvant,excipient, or vehicle with which the composition is administered. Acarrier is pharmaceutically acceptable after approval by a state orfederal regulatory agency or listing in the U.S. PharmacopeialConvention or other generally recognized sources for use in subjects.The pharmaceutical carriers include any and all physiologicallycompatible solvents, dispersion media, coatings, antibacterial andantifungal agents, isotonic and absorption delaying agents, and thelike. Examples of pharmaceutical carriers include, but are not limitedto, sterile liquids, such as water, oils and lipids such as, forexample, phospholipids and glycolipids. These sterile liquids include,but are not limited to, those derived from petroleum, animal, vegetableor synthetic origin such as, for example, peanut oil, soybean oil,mineral oil, sesame oil, and the like.

Suitable pharmaceutical excipients include, but are not limited to,starch, sugars, inert polymers, glucose, lactose, sucrose, gelatin,malt, rice, flour, chalk, silica gel, sodium stearate, glycerolmonostearate, talc, sodium chloride, dried skim milk, glycerol,propylene glycol, water, ethanol, and the like. In some embodiments, thecomposition can also contain minor amounts of wetting agents,emulsifying agents, pH buffering agents, or a combination thereof. Oralformulations, for example, can include standard carriers such as, forexample, pharmaceutical grades mannitol, lactose, starch, magnesiumstearate, sodium saccharine, cellulose, magnesium carbonate, and thelike. See Martin, E. W. Remington's Pharmaceutical Sciences.

As described herein, the compositions can take the form of lotions,creams, suspensions, emulsions, tablets, pills, capsules, powders,sustained-release formulations and the like. In some embodiments, thecompositions or formulations can be administered to a subject in anynon-parenteral manner known to one of skill whereas, in contrast, aparenteral administration involves piercing the skin or a mucousmembrane. Depending on the target tissue, the administration can betopical, oral, ocular, otologic, nasal, urogenital, rectal, dermal,vaginal or otherwise to a mucous membrane. Oral administration, forexample, can include digestive tract, buccal, and sublingualadministration, and a solid or liquid carrier can be used. One of skillwill appreciate that the therapeutic program selected, the agentsadministered, the condition of the subject, and the effects desired, canaffect the administration schedule and program used.

The compositions or formulations can be contained in forms that includetablets, troches, capsules, elixirs, beverages, suspensions, syrups,wafers, chewing gums, gels, hydrogels, and the like. Tablets, pills,capsules, troches liquids and the like may also contain binders,excipients, disintegrating agent, lubricants, glidants, chelatingagents, buffers, tonicity modifiers, surfactants, sweetening agents, andflavoring agents. Some examples of binders include microcrystallinecellulose, gum tragacanth or gelatin. Some examples of excipientsinclude starch or maltodextrin. Some examples of disintegrating agentsinclude alginic acid, corn starch and the like. Some examples oflubricants include magnesium stearate or potassium stearate. An exampleof a chelating agent is EDTA. Some examples of buffers are acetates,citrates or phosphates. Some examples of tonicity modifiers includesodium chloride and dextrose. Some examples of surfactants formicellation or increasing cell permeation include coconut soap, anionic,cationic or ethoxylate detergents. An example of a glidant is colloidalsilicon dioxide. Some examples of sweetening agents include sucrose,saccharin and the like. Some examples of flavoring agents includepeppermint, chamomile, orange flavoring and the like.

In the digestive tract, for example, a solid can include a pill,capsule, tablet, or time-release technology in some embodiments; and, aliquid can include a solution, soft gel, suspension, emulsion, syrup,elixir, tincture, or a hydrogel. Digestive tract administration caninclude oral or rectal administration using any method known to one ofskill. For buccal, sublingual, and sublabial administration, a solid caninclude an orally disintegrating tablet, a film, a lollipop, a lozenge,or chewing gum; and, a liquid can include a mouthwash, a toothpaste, anointment, or an oral spray.

One of skill understands that the amount of the agents administered canvary according to factors such as, for example, the type of disease,age, sex, and weight of the subject, as well as the method ofadministration. Dosage regimens may also be adjusted to optimize atherapeutic response. In some embodiments, a single bolus may beadministered; several divided doses may be administered over time; thedose may be proportionally reduced or increased; or, any combinationthereof, as indicated by the exigencies of the therapeutic situation andfactors known to one of skill in the art. It is to be noted that dosagevalues may vary with the severity of the condition to be alleviated, aswell as whether the administration is prophylactic, such that thecondition has not actually onset or produced symptoms. Dosage regimensmay be adjusted over time according to the individual need and theprofessional judgment of the person administering or supervising theadministration of the compositions, and any dosage ranges set forthherein are exemplary only and do not limit the dosage ranges that may beselected.

Articles of Manufacture

Kits that encompass finished, packaged and labelled products areprovided. These are articles of manufacture that include the appropriateunit dosage form in an appropriate vessel or container such as, forexample, a glass vial or other container that is hermetically sealed. Inthe case of dosage forms suitable for oral administration, the activeingredient, e.g. one or more agents including a dosage form taughtherein, may be suitable for administration topically, orally, rectally,vaginally, or the like. Alternatively, the unit dosage form may be asolid suitable for oral, transdermal, topical or mucosal delivery.

As with any such product, the packaging material and container aredesigned to protect the stability of the product during storage andshipment. In addition, the articles of manufacture can includeinstructions for use or other information material that can advise theuser such as, for example, a physician, technician or patient, regardinghow to properly administer the composition as a prophylactic,therapeutic, or ameliorative treatment of the disease of concern. Insome embodiments, instructions can indicate or suggest a dosing regimenthat includes, but is not limited to, actual doses and monitoringprocedures.

In some embodiments, the instructions can include informational materialindicating how to administer a composition for a particular use or rangeof uses, such as a particular indication taught herein, for example, aswell as how to monitor the subject for positive and/or negativeresponses to the administration.

In some embodiments, kits that contain a combination of topical, oral,rectal, or vaginal dosage forms for administrations to a subject arealso provided, as well as instructions for use in some embodiments, asthe kits can be designed for physicians, patients, or over the counteruse by any subject. In some embodiments, the kit is for protecting adermal tissue from UVA and UVB exposure, the kit comprising an extractof a Curcuma longa L. root in an oral dosage form; an extract of aCurcuma longa L. root in a topical dosage form; and, instructions foradministration of the topical dosage form, the oral dosage form, or acombination of the topical and oral dosage forms. In some embodiments,the kit is for treating any of the other indications taught herein, suchthat the kit can have any combination of topical, oral, and rectaldosage forms, and optionally instructions for use. The instructions canbe designed for the physician, the patient, or any subject, including,for example, instructions for mixing the components for administration,suggested dilution factors for various target sites, and potentialcombination therapies for combined administrations, such as topicalcombined with oral administration. The suggested dilution factors can beselected from the administration ranges taught herein, for example,which can be modified in some embodiments as desired, and incorporatedinto the compositions.

Without intending to be limited to any theory or mechanism of action,the following examples are provided to further illustrate the teachingspresented herein. It should be appreciated that there are severalvariations contemplated within the skill in the art, and that theexamples are not intended to be construed as providing limitations tothe claims.

Example 1 Extracting Curcuma longa L. Root with Ethanol

A variety of solvents may be used to extract the Curcuma longa L. root,and this example shows how the extraction can be done using ethanol.

Preparing Fresh Curcuma longa Root for Extraction

The extraction can be done with fresh Curcuma longa root, and the rootcan be prepared as follows:

Material

Fresh Curcuma longa root 50 g

Method

-   -   1. Wash the fresh root in water to remove any traces of soil or        other contaminants;    -   2. Drain to remove excess water;    -   3. Pat dry with paper towel;    -   4. Using a sharp knife or scapel, cut the root into pieces        approximately ½″×½″; and,    -   5. Use the moist, freshly-cut root pieces for extraction.

Preparing Dried Curcuma longa Root for Extraction

The Curcuma longa L. root may have been dried, for example, for storage.The dried root can be prepared for extraction as follows:

Material

Dried Curcuma longa root 30 g

Method

-   -   1. Wash the dried root in a dish of water to remove any traces        of soil or other contaminants, changing the wash water as        necessary;    -   2. Drain the wash water;    -   3. Add fresh water to cover the roots;    -   4. Soak overnight;    -   5. Drain the water and pat dry with paper towel;    -   6. Using a sharp knife or scapel, cut the root into pieces        approximately ½″×¹/2″;    -   7. Use the moist, freshly-cut root pieces for extraction;

Extracting with Ethanol

Ethanol is an efficient extraction solvent, and a process that can beused for extraction is as follows”.

Material

Prepared Curcuma longa root 50 g

Method

-   -   1. Transfer the prepared Curcuma longa root to a 250 mL beaker;    -   2. Pour 90% ethanol into the beaker until it covers the prepared        material and there is about 20 mL of ethanol above the level of        the prepared root;    -   3. Cover the beaker to prevent loss of ethanol;    -   4. Allow to macerate for 24 hours;    -   5. Decant the clear solvent;    -   6. Filter the remaining solvent under vacuum

Preparing a Dried Extract from the Ethanol Extract

The extracts can be removed from the ethanol and stored for use as a drycomposition. Moreover, ethanol can only be used in limited amountstopically and orally due to it's level of toxicity.

Material

Curcuma Longa Ethanol Extract.

Method

-   -   1. The prepared ethanol extract is transferred to the        round-bottomed flask of a rotary evaporator;    -   2. The solution is heated to a maximum temperature of 40° and        the condensed ethanol is collected via a condenser for the        purpose of re-use in subsequent extractions;    -   3. The resultant powder is stored in an amber bottle until        needed; and,    -   4. An alternate method is to add water to the ethanol extract        (for example, until the ethanol concentration is about 25%) and        then freeze drying the mixture.

Back-Extracting the Ethanol Extract into Oil

The extracts can be removed from ethanol, which is toxic and undesirableabove certain levels, by back-extracting the ethanol extract into asubstantially non-toxic oil, such as an edible oil.

Material

50 mL of an ethanol extract of Curcuma longa

Method

-   -   1. Transfer 50 mL of ethanol extract to 200 mL separating        funnel;    -   2. Add 50 mL of sesame oil;    -   3. Shake on a mechanical shaker for 30 minutes;    -   4. Store the separating funnel upright and allow the liquid        layers to separate for approximately 6 hours;    -   5. Carefully remove the sesame oil layer which can be used in        formulations of sunscreen; and,    -   6. One volume of ethanol may be extracted multiple times for        efficient extraction of the ethanol.

While the back extraction method, in principle, may be used to anethanol extract solution with any immiscible oil, the extraction appearsto be more effective when sesame oil or cotton seed oil are used whencompared to the use of liquid paraffin or glycerin. The relativeeffectiveness of the back-extraction using a particular oil can bemeasured and compared, for example, using a spectrophotometric analysis.

Preparing a Powdered Curcuma longa Root for Extraction

The Curcuma longa L. root can be converted into a powder. Extractionefficiency, for example, may be improved by powderizing the sample priorto extraction.

Material

Dried Curcuma longa Root

Method

-   -   1. Place about 50 gm of dried root pieces into a Wedgewood        mortar or other mortar with a rough surface (a glass mortar is        unsuitable).    -   2. With controlled tamping from a pestle, break up the pieces of        root into smaller pieces.    -   3. With firm pressure on the pestle, grind the material into a        coarse powder; and,    -   4. Using a coffee mill or similar laboratory grinder (such as a        hammer mill) grind the material into a fine powder

The fine Curcuma powder can be used for ethanol extraction or extractionwith oil as described herein, the advantage of the powder being a muchbetter yield of the sunscreen ingredient, as assessed by the UVspectrophotometric assay of the extract. Extracting the powder with aviscous oil, on the other hand, results in a viscous slurry which isdifficult to filter.

Example 2 Extracting Curcuma longa L. Root with Sesame Oil

The extracts removed using oils that are at least substantiallynon-toxic, as discussed herein, can be considered as highly favorable tothose in the art, as the extracts can be included directly into topicalor oral formulations without the use of additional separation steps thatinclude, for example, removal of the extract from the extraction solventprior to making an at least substantially non-toxic formulation for use.

Material

Prepared Curcuma longa root 50 g

Method

-   -   1. Transfer the prepared Curcuma longa root to a 250 mL beaker;    -   2. Pour sesame oil into the beaker until it covers the prepared        material;    -   3. Allow to macerate for 48 hours, with mixing after 24 hours;    -   4. Decant the clear solvent;    -   5. Add a second amount of sesame oil to cover the prepared        Curcuma longa root in the beaker;    -   6. Allow to macerate for a further 48 hours;    -   7. Decant the clear solvent;    -   8. Filter the remaining solvent under vacuum;

This type of extract appears to be a bit more dilute than the ethanolextract but can be incorporated directly into lotions and creams.Moreover, the concentration of the oil extract can be adjusted to apredetermined level by the addition of a calculated amount of dryextract of Curcuma longa after the spectrophotometric determination ofthe concentration of the oil.

Other oily solvents such as liquid paraffin, glycerin, squalene andcotton seed oil may be used although the more viscous solvents, such asliquid paraffin and glycerin, are more difficult to separate from theresidual matter.

Example 3 Assessing Extraction Efficiency of Different Solvent Systems

The extraction efficacy of different solvent systems were investigatedfor extraction of curcumin, deoxy curcumin, bis-deoxy curcumin andpossibly other curcumin derivatives and related substances (collectively“curcuminoids” hereafter) from plants of the family Zingerberaceae,especially Curcuma longa L. While not intending to be bound by anytheory or mechanism of action, it may be desired to utilize the wholeextract, since plant components are believed to work together tomodulate component activities. Biocompatible solvents were the focus,for at least the reasons set-forth herein, including the ability to usethe extract in the liquid form (including the extraction solvent)directly in pharmaceutical formulations that are at least substantiallynon-toxic and acceptable for human use, topically or orally.

Material

Liquid paraffin (light and heavy), glycerin, sesame oil, ethanol, cottonseed oil, polyethylene glycol (liquid grades including PEG 400),isopropyl myristate (IPM), polysorbate 80 (TWEEN 80) and combinations ofthese ingredients in 2 and 3 component mixtures with sesame oil, liquidparaffin, and cotton seed oil were tested.

Method

The method for determining the extraction efficiency of various solventswas as follows:

-   -   1. The results were obtained in triplicate and used to relate        the potency of the various extracts;    -   2. The ethanol extract as taught herein was used as a control,        the extract evaporated to dryness;    -   3. The ethanol extract was weighed as the dry extract and        dissolved in a specific volume of ethanol; and, from this first        solution, a series of dilutions of known concentration were made        as standard solutions to build a standard concentration curve to        compare results;    -   4. The absorption at the wavelength of maximum absorption (420        nm) was determined for each of the standard solutions and the        results were used to construct a standard curve of absorption        versus concentration;    -   5. Aliquots of experimental extracts, in various solvents, were        dissolved in ethanol and the absorption of the solution was        determined;

The results of extractions that were at least as efficient as theethanol extraction can be compared in the following table:

Amount Relative Extraction Extracted Efficiency (%) Solvent (g/100 mL)(Ethanol = 100%) Ethanol 6 100 IPM 6 100 TWEEN 80:IPM 6.6 110 (50:50mixture by volume) TWEEN 80:IPM 7 117 (20:80 mixture by volume) TWEEN 808.3 138

All extractions were compared to an ethanol extraction as a control, andit's worthy to reiterate that ethanol has limited utility since the dosefor internal consumption is limited and it cannot be used undiluted forrepeated application to large areas of the skin without side effects.

Otherwise ethanol, sesame oil, and cotton seed oil were found to bedesirable. Sesame oil and cotton seed oil, for example, were found toextract reasonable amounts of curcuminoids and could be incorporateddirectly into formulations such as emulsions for internal or externaluse. The IPM and TWEEN 80 (and their combinations) were found to be evenmore desirable, particularly were TWEEN 80 was used, having extractionefficiencies that were up to 38% better than the ethanol extraction. Ofthe different combinations of TWEEN 80 with IPM, namely 10%, 20%, 30%,and so on . . . up to 90% TWEEN 80 by volume, it was surprisingly foundthat a combination of either 20% or 50% TWEEN 80 appeared to give themost efficient extractions over other combinations, such as 30% and 40%TWEEN 80. We found, surprisingly, that TWEEN 80 (polysorbate 80 fromCroda Inc., Edison, N.J. 08837) was a better extraction solvent thangeneric polysorbate 80. TWEEN 80 HP performed even better than TWEEN 80(non-HP grade). This is surprising since the manufacturers only claim isthat the TWEEN 80 HP contains less impurities, such as peroxides, whichmay cause irritation at the application site. The higher purity grade isnot expected, on a theoretical basis, to have improved extractionability.

Example 4 Comparing Compositions of Ethanol Extracts to Those ProducedUsing IPM and TWEEN 80

This example compares the composition of an ethanol extract of Curcumalonga L. root to the TWEEN 80 extract and the IPM extracts taughtherein. The data compares (i) the total yield of curcuminoids and (ii)the relative proportions of individual curcuminoids.

Material

Prepared Curcuma longa Root

Method

-   -   1. prepare an ethanol extract as taught herein;    -   2. fractionate the crude extract;    -   3. isolate the individual components of the mixture;    -   4. identify the individual curcuminoids using NMR and LCMS and        compare to literature values; and,    -   5. Use TLC and HPLC to compare the ethanol extract with extracts        produced using TWEEN 80 and IPM extraction solvents.

Extraction Procedure

Turmeric root (3 g) was ground to a powder as taught herein andextracted overnight with 40 ml of ethanol as taught herein. The extractwas filtered through Whatman No. 1 filter paper, and the ethanol wasremoved using a rotary evaporator to give ˜180 mg of crude curcuminoids.The crude extract was kept in an amber glass bottle at room temperature.

Fractionation of Crude Extract

The curcuminoids were further fractionated using silica gel 60 columnchromatography with CH₂Cl₂/methanol gradient elution. The composition ofthe mobile phase was varied from 100%:0 to 90:10% to yield purefractions of Curcumin (1), Demethoxycurcumin (2), andBisdemethoxycurcumin (3), as follows:

Composition of Extract (%) TWEEN Compound R₁ R₂ MW MF EtOH 80 IPMCurcumin (1) OMe OMe 368.38 C₂₁H₂₀O₆ 71.32 73.82 70.94 Demethoxy- OMe H338.35 C₂₀H₁₈O₅ 18.06 17.05 18.66 curcumin (2) Bisdemethoxy- H H 308.33C₁₉H₁₆O₄ 10.62  9.13 10.4  curcumin (3)

All the fractions and eluates were monitored by thin layerchromatography (TLC) using pre-coated silica gel 60 F254, 0.25 mm glassbacked plates (Merck). The spots were detected by spraying with vanillin—H₂SO₄ reagent followed by heating. All chemicals and reagents used forTLC were of analytical grade. LC fractions that were tested and showedthe same pattern on TLC plates were pooled, and the organic solvent wasremoved to obtain the individual compounds in powder form foridentification and purity testing by additional standard chemical tests.

Identity and Purity Testing

The identity and purity of each curcuminoid was verified using TLC,high-performance liquid chromatography (HPLC)/mass spectrometry andNuclear Magnetic Resonance (NMR) analyses and by comparing to literaturedata. See, for example, Masuda, T et al. Phytochemistry 31: 3645-3647(1992); Syu, W.-J. et al. J. Nat. Prod. 61: 1531-1534 (1998); Huang,M.-T. et al. Carcinogenesis 16: 2493-2497 (1995); and, Byeoung-Soo Park,et al. J. Agric. Food Chem. 53: 9005-9009 (2005).

NMR

¹H- and ¹³C-NMR spectra were recorded on a BRUKER 600 AV IIIspectrometer with 5 mm TCI probe using tetramethylsilaneas an internalstandard. Chemical shifts are given in 6 (ppm).

Mass Spectrometry

Mass spectra were recorded on an AGILENT 1200 Series LC/MSD VL system.MS data were recorded on an Agilent Technology (HP) instrument with 5973Network Mass Selective Detector (MS model).

HPLC

A reverse phase HPLC method was developed using water:acetonitrilegradient 100:0 to 0:100% over 15 minutes. The purity and identity of thecurcuminoids was examined by reverse-phase HPLC(HITACHI, Elite LaChrome) equipped with a photodiode-array detector (L-2455) usingisolated pure compounds as a reference. The physico-chemical propertiesof these compounds are summarized below.

The structures of 1-3 were identified by comparing the spectral datawith those reported in the literature. See Id. The following evidencewas used for this comparison:

Curcumin [1]

Visual: Orange crystals;

LC-APCIMS (negative): MS, m/z 367; C₂₁H₂₀O₆;

HPLC: Retention time 12.41 min

1H NMR (CDCl3) data: δ 7.53 (2H, d, J=16 Hz, HC═C, 1,7-H, alkene,trans), 7.15 (2H, m, 6-ArH), 6.82 (4H, m, 2.5-ArH), 6.61 (2H, d, J=16,═CH—CO—, 2,6-H, alkene, trans), 5.91 (1H, s, CH═C(OH)—, 4-H), 3.89 (6H,s, 3-ArOCH₃), ppm;

13C NMR (CDCl₃) data: δ 185.0, 184.9, 150.6, 149.6, 142.3, 128.7, 124.3,122.44, 122.41, 116.7, 111.9, 102.2, 56.6 ppm.

Demethoxycurcumin [2]

Visual: Orange crystals;

LC-APCIMS (negative): MS, m/z 337; C₂₀H₁₈O₅; HPLC: Rt 12.20 min

1H NMR (CDCl3) data: δ 7.58 (d, 1H), 7.49 (d, 1H), 7.21 (s, 1H), 7.12(dd, 1H), 6.84 (d, 2H), 6.61 (t, 1H), 5.95 (br s, —OH), 3.89 (s, 3H,—OCH3)

13C NMR (CDCl₃) data: δ 185.1, 184.8, 161.3, 142.3, 142.0, 131.3, 128.7,124.3, 122.1, 117.0, 116.7, 111.9, 56.6 ppm.

Bisdemethoxycurcumin [3]

Visual: Orange crystals

LC-APCIMS (negative): MS, m/z 307; C₁₉H₁₆O₄; Rt 11.99 min

1H NMR (500 MHz, (CDCl₃) data: δ 7.57 (d, 2H), 7.49 (d, 4H), 6.82 (d,4H), 6.59 (d, 2H), 5.92 (s, 1H).

13C NMR (CDCl₃) data: δ 84.5, 161.3, 141.5, 131.3, 128.2, 122.1, 117.0,102.2.

TWEEN 80 and IPM extracts contain the same constituents as the ethanolextract and in approximately the same proportions. The identity andrelative proportions of curcuminoids in TWEEN 80 and IPM extracts weredetermined by HPLC.

Example 5 Yield and Stability of a Curcuma Longa L Root Extract

Yield: All of the examples taught herein provided a significant yield ofextract from the Curcuma longa L. root. When the Curcuma longa L. wasprepared as a powdered root, however, it provided larger yields of theextract from a single extraction. Extraction of 50 g of the powderedroot with ethanol, for example, produced about 1.5 gm of dried extractfrom a single extraction with 50 mL of ethanol. Extraction of thepowdered root with sesame oil, and alternatively squalene, also producedreasonable yields.

Stability: All extracts, stored in amber glass bottles, were stable formore than a year when tested by a spectrophotometric method.

Example 6 Formulations Produced Using IPM Extracts, and TWEEN 80: IPMExtracts

The extracts obtained using combinations of TWEEN 80 and IPM can bedirectly formulated from the liquid extract as an emulsion (creams orlotions) for external use; and, a limited amount of this mixture, or anemulsion produced from it, can be consumed by a human subject. Theextracts obtained using IPM are also capable of directly going intoformulation for topical or oral administration although, as noted, itcan be used with higher amounts of IPM when applied topically, forexample, as a sunscreen to the skin. This example provides someformulations for topical and oral use.

A simple oil formulation made directly from an extract obtained usingIPM only

IPM extract 75 g Sesame oil 25 g Butylated hydroxytoluene 0.01 g  Citrusextract 0.1 g 

-   -   1. Dissolve butylated hydroxytoluene in IPM extract with        stirring. Warm slightly to facilitate dissolution;    -   2. Add sesame oil and stir without further heating; and,    -   3. Add citrus extract and stir.

Another formulation . . .

A Lotion Formulation Made Directly from an Extract Obtained Using aCombination of 20% TWEEN 80 and 80% IPM

Extract with 20% Tween 80 in 80% IPM 50 g Span 80 4.86 g Water 45.14 gRose perfume 0.1 g

-   -   1. Gently warm the extract to 50° C.;    -   2. Add the SPAN 80 with continued warming and stir until        homogenous (the quantity of SPAN 80 was calculated as described        herein; IPM has a required HLB of 11.5);    -   3. Separately heat the water to 52° C.-54° C.;    -   4. Add the water all at once with rapid stirring and continue to        stir rapidly until emulsion is well formed;    -   5. Allow to cool with intermittent slow stirring; and,    -   6. When cooled almost to room temperature, add perfume and stir.

Another formulation . . .

A Cream Formulation Made Directly from an Extract Obtained Using aCombination of 20% TWEEN 80 and 80% IPM

Extract with 20% Tween 80 in IPM 50 g Polawax  4 g Water 46 g Lavenderperfume 0.1 g 

-   -   1. Gently warm the extract to 50° C.;    -   2. Add the Polawax with continued warming and stir until wax is        evenly dispersed;    -   3. Separately heat the water to 52° C.-54° C.;    -   4. Add the water all at once with rapid stirring and continue to        stir rapidly until emulsion is well formed;    -   5. Allow to cool with intermittent slow stirring; and,    -   6. When cooled almost to room temperature, add perfume and stir.

The extracts can be formulated into emulsions (e.g., for creams andlotions) that contain more than one oil phase ingredient as thefollowing example illustrates. The proportions HLB values of thedifferent oils incorporated are used to compute the overall required HLBfor the oil phase. The amount of a second emulgent to be added to theTWEEN 80 can then be easily determined. In the following example, whenliquid paraffin, sesame oil, and IPM (which have the required HLBs of12, 7 and 11.5, respectively) are combined in the proportions given inthis formulation, the overall required HLB is found to be 10.5. Theaddition of an emulgent blend, calculated on a percentage by weight ofthe total weight of emulgents, is 42% SPAN 80 (HLB=4.3) to 58% TWEEN 80(HLB=15) and results in a mixture with a desired HLB of 10.5.

Another formulation . . .

An Emulsion Made Directly from an Extract Obtained Using IPM Only, theEmulsion Formulated to Contain Multiple Oil Phase Components

Light liquid paraffin 20 g Sesame oil 10 g IPM 5 g Span 80 8.4 g Tween80 11.6 g Water 45 g Rose perfume 0.1 g

-   -   1. Gently warm the light liquid paraffin and sesame oil to 50°        C.;    -   2. Add the SPAN 80 and, with continued warming, stir until all        ingredients are evenly dispersed;    -   3. Add the TWEEN 80 extract and warm briefly;    -   4. Separately heat the water to 52° C.-54° C.;    -   5. Add the water all at once with rapid stirring and continue to        stir rapidly until emulsion is well formed;    -   6. Allow to cool with intermittent slow stirring; and,    -   7. When cooled almost to room temperature, add perfume and stir.

Another formulation . . .

A Microemulsion Made Directly from an Extract Obtained Using aCombination of 20% TWEEN 80 and 80% IPM

Extract with 20%:80% of Tween 80:IPM 50 g Isopropanol 20 g Water  4 g

-   -   1. Combine the extract and isopropanol while stirring (without        heating); and,    -   2. Add the water (no heat) with stirring.

Another formulation . . .

A Microemulsion Intended for External Use Made Directly from an ExtractObtained Using TWEEN 80

Extract with TWEEN 80 10 g IPM 40 g Isopropanol 20 g Water  4 g

-   -   1. Combine all ingredients with low intensity mixing and without        heating; and,    -   2. The microemulsion is observed as transparent, and the globule        size is measured using a dynamic light scattering with a Malvern        Nanosizer S and showed a single peak with a maximum of 262.4 nm.        The size, monodispersity, ease of formulation without use heat,        and clarity of the solution all confirm that it is a        microemulsion.

Another formulation . . .

An Edible Emulsion Made Directly from an Extract Obtained Using TWEEN 80

Extract with TWEEN 80 14.4 g  SPAN 80 5.6 g  Mineral Oil 20 g Water 60 g

-   -   1. Heat the mineral oil and SPAN to 50° C.-52° C. and stir        gently;    -   2. Add TWEEN extract and carefully heat to 50° C.-52° C., taking        care not to overheat;    -   3. Stir the mixture until uniform;    -   4. Heat the water to 52° C.-55° C., and add slowly but        all-at-once to the mixture; and,    -   5. Stir vigorously until the emulsion is well-formed, and then        stir occasionally until the emulsion reaches room temperature.

Another formulation . . .

An Edible NANOemulsion Intended to Increase Bioavailability UponConsumption, the Nanoemulsion Made Directly from an Extract ObtainedUsing TWEEN 80

Extract with TWEEN 80 14.4 g  SPAN 80 5.6 g  Mineral Oil 20 g Water 60 g

-   -   1. Heat the mineral oil and SPAN to 50° C.-52° C. and stir        gently;    -   2. Add TWEEN extract and carefully heat to 50° C.-52° C., taking        care not to overheat;    -   3. Stir the mixture until uniform;    -   4. Heat the water to 52° C.-55° C., and add slowly but        all-at-once to the mixture;    -   5. Stir vigorously until the emulsion is well-formed, and then        stir occasionally until the emulsion reaches room temperature;        and,    -   6. Pass the emulsion repeatedly through a microfluidizer to        reduce globule size to below 200 nm.

The microfluidizer can be sensitive to viscosity, and viscosity can beadjusted, for example by (i) adding water; (ii) decreasing the oilphase; or (iii) a combination thereof. The content of extract should bekept high, which can be done by keeping the TWEEN phase high in thisexample, as that's the source of the extract.

Non-aqueous liquids, such as extracts in TWEEN 80 or oils, or blends ofTWEEN 80 extracts with compatible absorption-enhancing liquids (such aspolyethyleneglycol 400), or mixtures of liquid extracts and moltensolids (such as GELUCIRE or solid polyethylene glycol, for example,polyethylene glycol 4000) may be filled into two piece gelatin capsulesand sealed to prevent leakage of the liquid fill from the capsules. Thesealing is performed using equipment such as the QUALICAPSHARD-CAPSULE-BAND-SEALING machine. The capsules, filled with theextract, are placed on slats appropriate for the size of capsule. Themachine draws the metal slat containing the capsules through the machineapplying a thin film of the sealing material (gelatin orhydroxypropylmethylcellulose (HPMC) solution, depending on whethergelatin or hydroxypropylmethylcellulose capsules are used). In a secondpassage through the machine, the seal is trimmed so that, effectively, anarrow band of sealing material is applied. Examples of the method ofpreparation of the sealing solutions are as follows:

Another example for preparing a dosage form . . .

Preparation of Gelatin Solution for Capsule Sealing

-   -   1. Measure 750 mL of ddH₂O (double-distilled water) into a clean        1 L beaker and heat the water to 57-60° C.;    -   2. Add the dye to the water, stir until the dye is completely        dissolved and a uniform solution is formed, and heat to 57-60°        C.;    -   3. Weigh 503 g of above water (57-60° C.) into a fresh 1 L        beaker (tared to zero);    -   4. Add 0.6 g of Tween-80 to the above beaker;    -   5. Weigh 141 g of gelatin into a tared weighing boat;    -   6. Add gelatin to the above beaker with hand stirring (spatula        or mixing rod);    -   7. Continue stirring until gelatin spreads evenly in the        solution. Cover the beaker with aluminum foil;    -   8. Place the beaker in an oven (preheated to 55° C.). Mix every        20-30 min, to prevent gelatin from sedimenting at the bottom;        and,    -   9. Continue mixing until all the gelatin is dissolved and a        clear uniform solution is formed (2-3 hr).

Another example for preparing a dosage form . . .

Preparation of HPMC Solution for Capsule Sealing

-   -   1. Weigh 100 g of HPMC powder into a fresh 1 L beaker (tared to        zero);    -   2. Weigh 344 g of absolute ethanol and add in increments to HPMC        powder with continuous hand mixing (use spatula or knife). Mix        for about 20-30 min until a smooth paste is formed (no clumps or        dry HPMC powder);    -   3. Add 181 gm of ddH₂O to above HPMC paste (HPMC+Alcohol) with        continuous hand mixing (no heating is required);    -   4. Add the dye to the water, stir until the dye is completely        dissolved and a uniform solution is formed; and,    -   5. Continue mixing until a clear solution is formed (15-20 min).        Once a clear solution is formed, cover the beaker with parafilm,        and let it stand for at least an hour prior to use (for air        bubbles to dissipate).

Another example for preparing a dosage form . . .

Suppository and Pessary Formulations

Suppository and pessary formulations for administration, respectively,to the rectum or vagina of a subject may be prepared by combining, forexample, the Tween 80 extract or the oil extract with a suitablesuppository or pessary base. In the molten state, the combined base andextract are poured into suitable molds which are available commercially.When solidified, the suppositories are removed from the mold. Examplesof suppository bases include Gelucire and polyethylene glycol. When thebase is available in various melting point grades, and is expected tomelt to release the active ingredient, a blend of grades may be usedsuch that the suppository or pessary melts at body temperature.Administration of the extracts by these routes and others, which do notinclude the swallowing of the dosage form, is expected to lead to higherabsorption of the drug since the degradation in the gastro intestinaltract and liver are avoided.

Examples of bases that could be used are gelatin, cocoa butter,polyethylene glycols, polyvinyl pyrrolidone, gelatin/glycerincombinations, esterfied fatty acids, polyoxyethelene sorbitans andpolyoxyethylene sorbitan fatty acid esters. Various additives may beincorporated including surfactants and absorption enhancers such asmedium chain (C8 to C12) fatty acids and fatty acid esters. Variouscommercial bases, which may contain mixtures of different components,are sold under the trade names Imhausen, Witepsol and Gelucire. Variousgrades of each of these are available for specific applications.Mixtures of various bases may also be utilized in order to obtain asuppository or pessary with the required properties. Other shapingmethods for forming the suppositories or pessaries including coldmolding and compression may also be used.

Example 7 Preparing a Sunscreen Lotion Containing Curcuma Longa L RootExtract

An emulsion of an extract of Curcuma longa L. can be produced in sesameoil using non-ionic surfactants, for example. A surfactant, orsurfactant pair, should first be chosen with the correct solubility forthe application. For non-ionic surfactants, we can calculate thehydrophilic-lipophilic balance (HLB) to match the HLB to theapplication.

Calculating Surfactant Ratio to Obtain Desired HLB

The HLB is the hydrophile-lipophile balance and gives an indication ofthe solubility of a surfactant.

The first step is to select a surfactant or surfactant pair. A desirablesurfactant or surfactant pair can be any surfactant or surfactant pairthought by one of skill to be useful in creating the emulsions that canbe used with the teachings provided herein. A SPAN surfactant and aTWEEN surfactant can be considered a desirable surfactant pair to createan emulsion of Curcuma longa L. extract in sesame oil. A SPANsurfactant, for example, can be referred to as a sorbitan alkanoate, andSPAN 60 has an HLB of 4.7. A TWEEN surfactant, for example, can bereferred to as an ethoxylated sorbitan alkanoate, and TWEEN 60 has anHLB of 14.9.

The following procedure can be used to calculate the relativepercentages of SPAN 60 and TWEEN 60 to be used in an emulsion withsesame oil to determine the ratio of surfactants to obtain the desiredHLB:

-   -   1. The desired HLB of sesame oil for oil-in-water emulsions has        been published as 7.0. See, for example, Haus, editor. Oral        Lipid-based Formulations: Enhancing the Bioavailability of        Poorly Water-Soluble Drugs. Informa Healthcare USA, Inc., New        York, N.Y. (2007).    -   2. If the fraction of SPAN 60=x,    -   3. Then, the fraction of TWEEN 60=1−x. Solving for x . . .

x(4.7)+(1−x)14.9=7; and,

x=0.775

As such, this means that a desired emulsion in this case should besuccessfully prepared using a mixture of 77.5% SPAN 60 and 22.5% TWEEN60 to obtain the 7.0 HLB for the mixture.

Making an Emulsion and Starting with a Sesame Oil Extract of Curcumalonga L.

An effective emulsion of a sesame oil extract can be made as follows:

Materials

-   -   1. Extract a Curcuma longa L. root as described herein;    -   2. Assume 200 g total weight of the emulsion;    -   3. Weigh 60 g of the sesame oil extract;    -   4. Assume 6% by wt of the surfactant pair, SPAN 60/TWEEN 60, HLB        7.0, and combine 9.3 g SPAN 60 with 2.7 g TWEEN 60 accordingly;        and,    -   5. Weigh 128 g water for the remainder.

Method

-   -   1. Carefully heat the sesame oil extract in a beaker to about        50° C., being careful not to overheat;    -   2. Add the SPAN 60 and stir with a glass rod to dissolve;    -   3. Separately, preheat the water to approximately 55° C.;    -   4. Add the TWEEN 60 to the water and stir with a glass rod to        dissolve;    -   5. Add the water with TWEEN 60 to the oil phase ingredients        while rapidly stirring the oil using a propeller stirrer. Stir        rapidly until emulsion forms, then stir occasionally until cool;        and,    -   6. Pack the lotion in an amber glass bottle.

The above is a basic formulation to which antioxidants, additionalemollients, vitamins, etc., may be added as desired. As an alternate wayof producing the sunscreen lotion, a dry extract of Curcuma longa may beincorporated into a lotion.

Making an Emulsion and Starting with a Dried Extract of Curcuma longa L.

Materials

Dried Curcuma longa extract as described herein.

Method

-   -   1. Weigh 25 mg of dried extract into a small, tared beaker;    -   2. Add approximately 100 g of sesame oil to the beaker;    -   3. Using a magnetic stirrer, stir the solution overnight until        all the powder is dissolved; and,    -   4. Use this oily solution to prepare an emulsion as described        herein.

Example 8 Novel Broad Spectrum Performance of Curcuma longa L. RootSuncreen as Compared to State of the Art Suncreens

The performance of current state-of-the-art sunscreen chemicals werecompared to that of the Curcuma longa L. extract in this example.

FIGS. 1A-1F compare the absorption spectra of representative currentstate-of-the-art sunscreen agents to the absorption spectra of Curcumalonga L., according to some embodiments. FIG. 1A shows the absorptionspectrum of octyl methoxycinnamate at a concentration of 5 ug/mlmethanol, FIG. 1B shows the absorption spectrum of octocrylene at aconcentration of 0.5 ug/ml methanol, FIG. 10 shows the absorptionspectrum of oxybenzone at a concentration of 1 ug/ml methanol, FIG. 1Dshows the absorption spectrum of homosalate at a concentration of 20ug/ml methanol, and FIG. 1E shows the absorption spectrum of avobenzoneat a concentration of 5 ug/ml methanol.

The representative sunscreen agents show peaks in the absorptionspectrum between the wavelengths of 290 nm to 400 nm which is the rangeof interest. Ideally, a desired agent would absorb energy across theentire range of 290 nm to 400 nm. In FIGS. 1A-1E, for example, a varietyof peak positions are seen in the representative agents, where a high(desirable) absorption can be shown at the wavelength of the peak butthere is usually a low or zero absorption at the trough, which is, ofcourse, undesirable. Even combinations of sunscreen agents with peaksand troughs may leave areas within the wavelength range that are notcovered, leaving wavelengths for which there is little or no UVprotection, an undesirable situation.

FIG. 1A shows that octyl methoxycinnamate has practically no absorptionat wavelengths higher than approximately 350 nm. FIG. 1B shows thatoctocrylene has an absorption that is very low above about 370 nm and ispractically zero beyond about 380 nm. FIG. 1C shows that oxybenzone hasan absorption that is practically zero above about 380 nm. FIG. 1D showsthat homosalate has an absorption that is practically zero above about340 nm. FIG. 1E shows that only avobenzone has a broad spectrum ofactivity but it, too, has low absorption beyond about 390 nm.

FIG. 1F, on the other hand, shows the very broad spectrum of absorptionof the Curcuma longa L. extract, according to some embodiments. The scanof the extract of Curcuma longa shows no dips or troughs in theabsorption of UV light from the wavelength of 280 nm to 400 nm. Inessence, it was surprising and unexpected to find that Curcuma longa L.extract provided such a broad range of protection from the harmful raysof the sun across the entire wavelength range for which humans needsunscreen protection.

As seen from FIG. 1F, the UV absorption spectrum extends across theentire range from 280 nm to 400 nm. Moreover, unlike the other agentstested, the level of absorption does not decrease as it approaches thehigher UV range but, rather, it surprisingly increases. The UVabsorption of the Curcuma longa L., for example, covers all of theimportant UVA region and, moreover, the level of UVA₁ absorption is highrather than minimal at best, as with state-of-the-art sunscreens. Assuch, the Curcuma longa L. is a much superior sunscreen agent whencompared to the representative sunscreen agents using at least thecriteria of the Jun. 17, 2011, the FDA report discussed herein. Restatedin this section for emphasis, the FDA stated that

(i) sunscreen products should have both UVA and UVB protection; and,

(ii) the UVA range has two unique areas of interest:

-   -   the UVA₁ area ranging from 340 nm to 400 nm; and,    -   the UVA₂. area ranging from 320 nm to 340 nm.

The FDA also stated that too much emphasis has been placed on UVB, thatat least 20% of the protection should be in the UVA₂ region, and atleast 60% of the protection should be in the UVA₁ region, stressing therelative importance of UVA₁ region. Accordingly, the UV absorptionspectrum of the extract of Curcuma longa is not only unique, but alsohighly desirable when compared to the state-of-the-art sunscreen agentscurrently in use.

The performance of the extract as a sunscreen was quite surprising,particularly due to the broad spectrum of protection from the sunwithout requiring use of chemical additives such as TiO₂ to screen bothUVA and UVB. It should be appreciated that the very broad UV absorptionpeak of the present extract, covering UVA₁, UVA₂ and UVB, is unique inthe art and offers better protection than multiple compounds that cannotcover this broad UV wavelength range. One of skill will appreciate that,even when multiple sunscreen agents are used, each of the agents candemonstrate its own peak across the absorption spectrum, the resultbeing that the combined agents still contain areas of poor UVabsorption.

By the addition of small amounts of blue and red color, the naturalcolor of the extract can be made into a tan color. When applied to theskin of a darker person, this color blends with the skin color and hasno further impact on appearance. However, when applied to a fair-skinnedperson, it will produce a “tanned” appearance. It is possible for oneskilled in the art, to produce a range of shades acceptable tofair-skinned persons of different skin tones which, simultaneously,protects from the UV radiation of the sun and offers a tannedappearance. It is also possible for one skilled in the art to add anatural plant extract which has a blue/purple color (such as blueberryextract) to attain the same skin tone effect. The advantage of thelatter method is that the chosen natural extract may add additionalvalue to the product, such as, in the case of blueberry extract, asecond source of antioxidants which complement the antioxidant effect ofthe curcumin extract.

Example 9 Method of Testing Sunscreen Formulations on Human Subjects

The methods provided in this section are in accord with the methodsdetailed in the FDA's final rule on sunscreen preparations and testingpublished in 21 CFR 352.70(a), dated 17 Jun. 2011 (“regulations”hereafter).

Subjects

Thirteen (13) subjects can be enrolled with the objective of having atleast 10 subjects complete the study. Compared to systemically-absorbeddrug evaluation studies which, often, involve some drug side effects,the dropout rate for this study is not expected to be high. Thereferenced FDA guidelines state that at least 10 subjects are needed forvalid results.

Exclusion Criteria

Exclusion criteria include the following:

-   -   1. Previous history of extreme sensitivity to sunlight with        severe sunburn occurring with short exposure; or abnormal        responses to sunlight, such as a phototoxic or photoallergic        response with normal exposure;    -   2. Taking medication (topical or systemic) that is known to        produce abnormal reactions to light;    -   3. Uneven skin tones on the areas of the back to be tested;    -   4. Any lesions on the area of exposure;    -   5. Tattoos or scarring on the area of exposure;    -   6. Diabetics; and,    -   7. Pregnant women.

Inclusion Criteria

Male and female subjects who are fair skinned shall be selected based ontheir classification, using the following guidelines, as skin types I,II, and III. The classification depends on the response of the fairskinned people to the first 30 to 45 minutes of sun exposure after awinter season of no sun exposure:

-   -   1. Always burns easily; never tans (sensitive);    -   2. Always burns easily; tans minimally (sensitive);    -   3. Burns moderately; tans gradually (light brown) (normal);    -   4. Burns minimally; always tans well (moderate brown) (normal);    -   5. Rarely burns; tans profusely (dark brown) (insensitive); and,    -   6. Never burns; deeply pigmented (insensitive).

Subject Selection

Subjects will be selected from a pool of volunteers, each of whichprovide legally effective written informed consent. A medical historywill be taken from all subjects with emphasis on the effects of sunlighton their skin. The general health of the individual will be ascertainedby means of a physical examination. In addition, the test site will beinspected with emphasis on exclusion criteria 3, 4, 5. The presence ofnevi, blemishes, or moles will be acceptable if, in the physician'sjudgment, they will not interfere with the study results. Excess hair onthe back is acceptable if the hair is clipped. Shaving is unacceptablesince too much epidermis may be removed.

Trial Materials

The trial materials will consist of:

-   -   1. TEST MATERIAL—the sunscreen cream or lotion containing the        Curcuma longa L. extract as the sun screen material (TEST); and,    -   2. REFERENCE STANDARD—the standard consists of a cream        containing 7% padimate O and 3% oxybenzone as the sunscreen        materials. (REFERENCE). The composition of the reference        standard is given as follows in the regulations*:

Ingredients % by weight Part A Lanolin 4.50 Cocoa butter 2.00 Glycerylmonostearate 3.00 Stearic acid 2.00 Padimate O 7.00 Oxybenzone 3.00 PartB Purified water USP 71.60 Sorbitol solution 5.00 Triethanolamine 99percent 1.00 Methylparaben 0.30 Propylparaben 0.10 Part C Benzyl alcohol0.50 Part D Purified water USP QS to 100 g *Federal Register/Vol. 76,No. 117/Friday, Jun. 17, 2011: Labeling and Effectiveness Testing;Sunscreen Drug Products for Over-the-Counter Human Use.

Method of Making REFERENCE

The method can be performed as follows:

-   1. Add the ingredients of Part A into a suitable stainless steel    kettle equipped with a propeller agitator. Mix at 77 to 82° C. until    uniform;-   2. Add the water of Part B into a suitable stainless steel kettle    equipped with a propeller agitator and begin mixing at 77 to 82° C.    Add the remaining ingredients of Part B and mix until uniform;-   3. Add the batch of 1 to the batch of 2 and mix at 77 to 82° C.    until smooth and uniform. Slowly cool the batch to 49 to 54° C.;-   4. Add the benzyl alcohol of Part C to the batch of 3 at 49 to    54° C. Mix until uniform. Continue to cool batch to 35 to 41° C.;-   5. Add sufficient water of Part D to the batch of 4 at 35 to 41° C.    to obtain 100 grams of SPF standard. Mix until uniform. Cool batch    to 27 to 32° C.

UV Light Source for Human Testing

This study will use a solar simulator that conforms to the regulations.The simulator should be capable of providing UV radiation such that:

-   1. UVA II (320-340 nm) should equal or exceed 20 percent of the    total UV (290 nm-400 nm) irradiance;-   2. UVA I (340 nm-400 nm) should equal or exceed 60 percent of the    total UV (290 nm-400 nm) irradiance;-   3. The total irradiance limit should be 1500 W/m²; and,-   4. The total irradiance range should be 250-1400 nm.

Method of Human Testing

The method can be performed as follows:

-   1. The test location will be on the back of the subject between the    beltline and the shoulder blade (scapulae) and lateral to the    midline. This is the area where the subject's natural sensitivity to    UV radiation will be determined, the sunscreen and standard will be    applied and assessed;-   2. Assessment of the standard and test will be based on the    principle of minimal erythemal dose (MED) i.e. the lowest dose of UV    radiation that will cause erythema. Firstly, MED will be assessed on    the patient's skin with no application of any agent (unprotected) in    order to obtain his base-line or control MED. Thereafter, the test    formulation and the standard formulation will be applied and the MED    determined again.-   3. Within the test location, each area for applying a product or the    standard sunscreen (“test site”) will be outlined with ink. Each    test site will be a minimum of 30 cm² in area. If the person is to    be tested in an upright position, the lines will be drawn on the    skin with the subject upright. If the subject is to be tested while    prone, the markings will be made with the subject prone. Thus    stretching and wrinkling of the skin will not alter the extent of    the area to which TEST and STANDARD are applied.-   4. Each test site area will be further divided into four or five    subsites that each have an area of at least 0.5 cm². The distance    between subsites is at least 0.8 cm.-   5. Both the test sunscreen product and the standard sunscreen are    applied to their respective test areas at a standardized rate. The    rate specified in the regulation is 2 milligrams per square    centimeter. The density of the formulations will first be determined    and the standard weight will be added volumetrically using a    suitably sized syringe.-   6. Sunscreen drug products and the test product will be evaluated in    the same study. These products will be applied in a blinded,    randomized manner. Each product will be spread using a finger cot.    Presaturation of finger cot is not required.-   7. After applying a product, a waiting period of at least 15 minutes    is required before exposing the test site areas to radiation.-   8. Next, each test subsite within a test site area is be subjected    to a specified dosage of UV radiation. A series increasing UV    radiation exposures is used for the determination of the MED.-   9. In order to maintain the blind, the person who evaluates the MED    responses is not the same person who applied the sunscreen drug    product to the test site or administered the doses of UV. In    addition, care is taken that the evaluator does not know this    information.-   10. After UV radiation exposure from the solar simulator is    completed, the subject's immediate responses to the exposure are    recorded. It is important to include all responses. Examples of    immediate responses include the following:    -   a. darkening or tanning, typically greyish or purplish in color,        fading in 30 to 60 minutes, and attributed to photo-oxidation of        existing melanin granules;    -   b. immediate reddening, fading rapidly, and viewed as a normal        response of capillaries and venules to heat, visible and        infrared radiation; and,    -   c. an immediate generalized heat response, resembling prickly        heat rash, fading in 30 to 60 minutes, and apparently caused by        heat and moisture generally irritating to the skin's surface.-   11. After the immediate responses are noted, shield the exposed area    of each subject from further UV radiation for the remainder of the    test day.-   12. The MED is determined 22 to 24 hours after exposure and the    determination is made under the following conditions:-   13. The source of illumination is either a tungsten light bulb or a    warm white fluorescent light bulb that provides a level of    illumination of 450 to 550 lux at the test site-   14. The subject should be in the same position used when the test    site was irradiated.-   15. Determine the smallest dose of energy, for each series of    exposures, which produces redness. To be counted, the redness must    reach the borders of the exposure site at 22 to 24 hours post    exposure. Somewhat more intense erythemas must also be produced, in    order to know the minimal dose that produced erythema. The goal is    to have some exposures that produce absolutely no effect, some that    have redness reaching the borders of the exposure site (and can    therefore be counted as minimal exposure producing the required    effect) and some exposures that produce a greater effect than is    needed (which will not be counted).-   16. Reject the test data if the exposure series fails to elicit an    MED response on either the treated or unprotected skin sites, or if    the responses on the treated sites are randomly absent (which    indicates the product was not spread evenly).

Definitions for Human Testing

-   MED—minimal erythema dose—The MED is the smallest UV dose that    produces perceptible redness of the skin (erythema) with clearly    defined borders at 16 to 24 hours after UV exposure.-   MEDu—the MED for unprotected skin—This is determined on a test site    that does not have sunscreen applied.-   MEDp—the MED for protected skin—This is determined on a test site    that has sunscreen applied-   ssMEDp—the MEDp for the sunscreen standard-   tpMEDp—the MEDp for the sunscreen test product-   Evaluation of test subsites—In order that the person who evaluates    the test subsites is not biased, he/she should not be the same    person who applied the sunscreen drug product to the test site or    administered the UV doses.-   Immediate Responses—After UV doses are administered, all immediate    responses should be recorded. These may include an immediate    darkening or tanning, typically grayish or purplish in color, which    fades in 30 to 60 minutes; an immediate reddening at the subsite,    due to heating of the skin, which fades rapidly; and an immediate    generalized heat response, spreading beyond the subsite, which fades    in 30 to 60 minutes-   MED Determination—After the immediate responses are noted, each    subject should shield the exposed area from further UV radiation    until the MED is determined. This must be determined 16 to 24 hours    after UV exposure i.e. the final MEDu, ssMEDp, and tpMEDp are    typically determined the day following determination of the initial    MEDu. Evaluate the erythema responses of each test subsite using    either tungsten or warm white fluorescent lighting that provides at    least 450 lux of illumination at the test site. For the evaluation,    the test subject should be in the same position as he or she was    when the test site was irradiated.

SPF Determination—

-   1. Calculate an SPF value for each test subject or individual (SPFi)    as follows:

${SPFi} = \frac{MEDp}{MEDu}$

-   2. Calculate the MEAN SPF and the standard deviation (s) from the    SPFi values. Calculate the standard error (SE), which equals s/√n    (where n equals the number of subjects who provided valid test    results). Obtain the t value from Student's t distribution table    corresponding to the upper 5-percent point with n−1 degrees of    freedom. Determine the labeled SPF value, which equals the largest    whole number less than

MEAN SPF−(t*SE)

-   3. In order for the SPF determination of a test product to be    considered valid, the SPF value of the SPF standard should fall    within the standard deviation range of the expected SPF (i.e.,    16.3±3.43).

Example 10 Testing the Anti-Oxidant Activity of the Curcuma Longa L.Extract

The antioxidant activity of Curcuma longa L. was assessed and comparedto resveratrol, a well-known antioxidant.

Material

A TWEEN extract of Curcuma longa produced as taught herein;

Resveratrol (trans-3,5,4′-trihydroxystilbene), is an extract of the skinof red grapes and found in red wines.

Method

The comparisons were done using the DPPH (1,1-diphenyl-2-picrylhydrazyl)assay, a well-established method for testing antioxidant activity. TheDPPH assay is based on the inhibition by antioxidants of the absorbanceof the DPPH free radicals, having a characteristic absorption spectrumshowing a maximum at 517 nm.

The resveratrol is dissolved in methanol, and both the TWEEN extract andresveratrol were tested at the same concentration of 4.2 mg/mL.

A 50 μM DPPH-free radical solution was prepared by dissolving the1,1-diphenyl-2-picrylhydrazyl in methanol.

-   -   1. The absorbance of DPPH-free radical solution (prepared as        above) was read at 517 nm (control);    -   2. 0.1 mL of Tween extract of Curcuma longa was diluted with 0.4        mL of methanol;    -   3. 2 mL of DPPH reagent was added and its absorbance was read        after allowing 60 seconds reaction time, using 0.1 mL of Tween        diluted with methanol as a blank;    -   4. The absorbance of solution 3 was read at 60-second intervals        using an automated function on the spectrophotometer;    -   5. A solution of resveratrol (4.2 mg/mL) was prepared;    -   6. 0.1 mL of the resveratrol solution was diluted with 0.4 mL of        methanol and 2 mL of DPPH reagent was added and the absorbance        of this solution was read after allowing 60 seconds reaction        time, using methanol as a blank;    -   7. The absorbance of solution 6 was read at 60-second intervals        using an automated function on the spectrophotometer; and,    -   8. The % inhibition of DPPH-free radicals was calculated using        the following equation, where A is absorbance, the control is        DPPH, and the samples are the TWEEN extract and the resveratrol:

% inhibition=[A _(control) −A _(sample))/A _(control)]×100

FIGS. 2A and 2B compare the antioxidant activity of the TWEEN extract ofCurcumin longa to the antioxidant activity of resveratrol, according tosome embodiments. FIG. 2A shows the comparison 200 over a time frame of20 minutes, where it can be seen that the antioxidant activity of theTWEEN extract 205 and the resveratrol 210 is comparable at timesexceeding 5 minutes. FIG. 2B shows the comparison over the time frame of5 minutes, where it is clear that the activity of TWEEN extract 205 ismultiples higher than the resveratrol 210 for the first couple ofminutes, and the activities for the TWEEN extract 205 and theresveratrol 210 become comparable at times ranging from about 5 minutesto about 20 minutes.

As such, it can be concluded that the antioxidant activity of the TWEENextract is comparable to a well-known antioxidant having an accepted anddesirable activity that provides accepted and desirable therapeuticresults. Resveratrol has shown to be promising at preventing and/ortreating at least the following:

-   -   1. cancer in animals;    -   2. reducing the risk of heart disease by inhibition of vascular        cell adhesion, inhibition of vascular smooth muscle cell        proliferation, inhibition of platelet aggregation, inhibition of        LDL peroxidation, stimulation of endolethelial nitric oxide        synthase (eNOS) activity;    -   3. diabetes, through it's hypoglycemic and hypolipidemic        effects, and it also ameliorates common diabetes symptoms, such        as polyphagia, polydipsia, and body weight loss;    -   4. neuroprotective effects, including reducing plaque formation        in brains, a component of Alzheimer's disease and other        neurodegenerative diseases    -   5. anti-inflammatory effects, including inhibiting both acute        and chronic phases of the inflammatory process, and showing        promise as a potential therapy for arthritis, for example, the        progression of inflammatory arthritis.    -   6. antiviral effects, including inhibiting herpes simplex virus        (HSV) types 1 and 2 replication by inhibition of an early step        in the virus replication cycle, inhibiting HSV replication in        the vagina, limiting extravaginal disease, as well as inhibiting        varicella-zoster virus, influenza viruses, respiratory viruses,        and human cytomegalovirus. It's also has shown synergistic        enhancements in the treatment of the anti-HIV-1 activity of        several anti-HIV drugs.    -   7. Testosterone level, including increasing hormone level and        sperm count.

As such, given the comparative antioxidant activity provided in thisexample, in addition to the literature on Curcuma longa, this suggeststhat the extracts of Curcumin longa may be promising in treating atleast these conditions as well.

We claim:
 1. A method of preparing a pharmaceutically acceptable liquidextract of Curcuma longa L, the method comprising: macerating at least aportion of a Curcuma longa L. root for an effective time in the at leastsubstantially non-toxic extraction solvent that is miscible withphenolic diketones that include curcumin, demethoxycurcumin, andbisdemethoxycurcumin; and, pharmaceutically acceptable as a carrier forthe phenolic diketones, the carrier having (i) an acute oral toxicitywith an LD50 of at least 49,700 mg/kg or (ii) an acute dermal toxicityof at least 5000 mg/kg; wherein, the macerating includes covering theportion of the root with the extraction solvent; and, separating theextraction solvent from the macerated root to create a liquid extract ofCurcuma longa L.
 2. The method of claim 1, wherein the extractionsolvent comprises TWEEN 80 HP.
 3. The method of claim 1, wherein theextraction solvent comprises isopropyl myristate.
 4. The method of claim1, wherein the emulsifying includes adding a pharmaceutically acceptableoil to the liquid extract to create the emulsion of the liquid extract.5. The method of claim 1, wherein the emulsifying includes adding anemulgent to the liquid extract to create the emulsion of the liquidextract.
 6. The method of claim 4, wherein the pharmaceuticallyacceptable oil comprises sesame oil.
 7. The method of claim 4, whereinthe pharmaceutically acceptable oil comprises cotton seed oil.
 8. Themethod of claim 1 further comprising placing the purified liquid extractinto a dosage form for treating an inflammation of a tissue of asubject.
 9. The method of claim 8, wherein the tissue is a dermaltissue.
 10. The method of claim 8, wherein the tissue is a mucosaltissue.
 11. The method of claim 8, wherein the tissue isgastrointestinal tissue.
 12. The method of claim 1 further comprisingplacing the purified liquid extract into a dosage form for treating awounded tissue of a subject.
 13. The method of claim 12, wherein thetissue is a dermal tissue.
 14. The method of claim 12, wherein thetissue is a mucosal tissue.
 15. The method of claim 12, wherein thetissue is gastrointestinal tissue.
 16. A method of preparing apharmaceutically acceptable emulsion of a liquid extract of Curcumalonga L, the method comprising: macerating at least a portion of aCurcuma longa L. root for an effective time in the at leastsubstantially non-toxic extraction solvent that is miscible withphenolic diketones that include curcumin, demethoxycurcumin, andbisdemethoxycurcumin; and, pharmaceutically acceptable as a carrier forthe phenolic diketones, the carrier having (i) an acute oral toxicitywith an LD50 of at least 49,700 mg/kg or (ii) an acute dermal toxicityof at least 5000 mg/kg; wherein, the macerating includes covering theportion of the root with the extraction solvent; separating theextraction solvent from the portion of the macerated root to create aliquid extract of Curcuma longa L.; and, emulsifying the liquid extract.17. The method of claim 16, wherein the extraction solvent comprisesTWEEN 80 HP.
 18. The method of claim 16, wherein the extraction solventcomprises isopropyl myristate.
 19. The method of claim 16, wherein theemulsifying includes adding a pharmaceutically acceptable oil to theliquid extract to create the emulsion of the liquid extract.
 20. Themethod of claim 16, wherein the emulsifying includes adding an emulgentto the liquid extract to create the emulsion of the liquid extract. 21.The method of claim 19, wherein the pharmaceutically acceptable oilcomprises sesame oil.
 22. The method of claim 19, wherein thepharmaceutically acceptable oil comprises cotton seed oil.
 23. Themethod of claim 16 further comprising placing the purified liquidextract into a dosage form for treating an inflammation of a tissue of asubject.
 24. The method of claim 23, wherein the tissue is a dermaltissue.
 25. The method of claim 23, wherein the tissue is a mucosaltissue.
 26. The method of claim 23, wherein the tissue isgastrointestinal tissue.
 27. The method of claim 16 further comprisingplacing the purified liquid extract into a dosage form for treating awounded tissue of a subject.
 28. The method of claim 27, wherein thetissue is a dermal tissue.
 29. The method of claim 27, wherein thetissue is a mucosal tissue.
 30. The method of claim 27, wherein thetissue is gastrointestinal tissue.